Anti-cgrp compositions and use thereof

ABSTRACT

The present invention is directed to antibodies and fragments thereof having binding specificity for CGRP. Another embodiment of this invention relates to the antibodies described herein, and binding fragments thereof, comprising the sequences of the V H , V L  and CDR polypeptides described herein, and the polynucleotides encoding them. The invention also contemplates conjugates of anti-CGRP antibodies and binding fragments thereof conjugated to one or more functional or detectable moieties. The invention also contemplates methods of making said anti-CGRP antibodies and binding fragments thereof. Embodiments of the invention also pertain to the use of anti-CGRP antibodies, and binding fragments thereof, for the diagnosis, assessment and treatment of diseases and disorders associated with CGRP.

This application claims the benefit of U.S. Provisional Application No.61/488,660 (Atty. Docket No. 67858.730300) filed May 20, 2011, entitled“ANTI-CGRP COMPOSITIONS AND USE THEREOF” which is hereby incorporated byreference in its entirety.

The instant application contains a Sequence Listing which has beensubmitted in ASCII format via EFS-Web and is hereby incorporated byreference in its entirety. Said ASCII copy, created on May 18, 2012, isnamed 67858o730301.txt and is 203,818 bytes in size.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention pertains to antibodies and fragments thereof (includingFab fragments) having binding specificity to human Calcitonin GeneRelated Peptide (hereinafter “CGRP”). The invention also pertains tomethods of screening for diseases and disorders associated with CGRP,and methods of preventing or treating diseases and disorders associatedwith CGRP by administering said antibodies or fragments thereof.

2. Description of Related Art

Calcitonin Gene Related Peptide (CGRP) is produced as a multifunctionalneuropeptide of 37 amino acids in length. Two forms of CGRP, theCGRP-alpha and CGRP-beta forms, exist in humans and have similaractivities. CGRP-alpha and CGRP-beta differ by three amino acids inhumans, and are derived from different genes. The CGRP family ofpeptides includes amylin, adrenomedullin, and calcitonin, although eachhas distinct receptors and biological activities. Doods, H., Curr. Op.Invest. Drugs, 2(9):1261-68 (2001). CGRP is released from numeroustissues such as trigeminal nerves, which when activated releaseneuropeptides within the meninges, mediating neurogenic inflammationthat is characterized by vasodilation, vessel leakage, and mast-celldegradation. Durham, P. L., New Eng. J. Med., 350 (11):1073-75 (2004).The biological effects of CGRP are mediated via the CGRP receptor(CGRP-R), which consists of a seven-transmembrane component, inconjunction with receptor-associated membrane protein (RAMP). CGRP-Rfurther requires the activity of the receptor component protein (RCP),which is essential for an efficient coupling to adenylate cyclasethrough G proteins and the production of cAMP. Doods, H., Curr. Op.Invest. Drugs, 2(9):1261-68 (2001).

Migraines are neurovascular disorder affecting approximately 10% of theadult population in the U.S., and are typically accompanied by intenseheadaches. Approximately 20-30% of migraine sufferers experience aura,comprising focal neurological phenomena that precede and/or accompanythe event. CGRP is believe to play a prominent role in the developmentof migraines. For example, plasma concentrations of CGRP were identifiedelevated in jugular venous blood during the headache phase of migraines,to the exclusion of other neuropeptides. Moreover, according toArulmozhi et al, the following has been identified in migrainesufferers: (1) a strong correlation between plasma CGRP concentrationsand migraines; (2) the infusion of CGRP produced a migraine-likeheadache; (3) baseline CGRP levels were elevated; and (4) changes inplasma CGRP levels during migraine attacks significantly correlated withheadache intensity. Arulmozhi, D. K., et al., Vas. Pharma., 43: 176-187(2005).

One effective treatment for migraines is the administration of triptans,which are a family of tryptamine-based drugs, including sumatriptan andrizatriptan. Members of this family have an affinity for multipleserotonin receptors, including 5-HT_(1B), 5-HT_(1D), and 5-HT_(1F).Members of this family of drugs selectively constrict cerebral vessels,but also cause vasoconstrictive effects on coronary vessels. Durham, P.L., New Eng. J. Med., 350 (11):1073-75 (2004). There is a theoreticalrisk of coronary spasm in patients with established heart diseasefollowing administration, and cardiac events after taking triptans mayrarely occur. Noted to be contraindicated for patients with coronaryvascular disease.

Similarly, pain may often be addressed through the administration ofcertain narcotics or non-steroidal anti-inflammatory drugs (NSAIDs).However, the administration of these treatments may occur at the cost ofcertain negative consequences. NSAIDs have the potential to cause kidneyfailure, intestinal bleeding, and liver dysfunction. Narcotics have thepotential to cause nausea, vomiting, impaired mental functioning, andaddiction. Therefore, it is desirable to identify alternative treatmentsfor pain in order to avoid certain of these negative consequences.

CGRP is believed to play a role in a multitude of diseases anddisorders, including but not limited to migraines, headaches, and pain.

For example, CGRP reportedly may correlate to or even pay a causal playa role in overactive bladder. Evidence that CGRP may correlate tooveractive bladder condition includes the fact that CGRP is present inurinary tract, DRG and spinal cord—(Wharton et al., 1986 Neurosci(3):727) and also that C-fiber afferents are critical for carryingimpulses involved in micturition to spinal cord (Yoshida et al., 2011 JPharmacol Sci (112):128). Further, it has been reported that theintravesical administration of Botox suppresses CGRP and significantlyreduces intercontraction interval in acetic acid—induced bladder painmodel (Chuang et al., 2004 J Urol (172):1529; Chuang et al., 2009 J Urol(182):786)

Evidence that CGRP may play a causal role in this condition is a recentpublished patent application containing data purportedly suggesting thatan anti-CGRP Ab disclosed therein reduced the number of bladdercontractions in a turpentine-oil—induced overactive bladdermodel—(Pfizer WO 2011/024113)).

Due to the perceived involvement of CGRP in these and other disorders,there remains a need in the art for compositions and methods useful forpreventing or treating diseases and disorders associated with CGRP,while avoiding adverse side effects. There remains a need in the art forcompositions or methods that reduce or inhibit diseases or disordersassociated with CGRP, such as migraines, headaches, overactive bladder,and pain.

BRIEF SUMMARY OF THE INVENTION

The present invention is directed to specific antibodies and fragmentsthereof having binding specificity for CGRP, in particular antibodieshaving desired epitopic specificity, high affinity or avidity and/orfunctional properties. Another embodiment of this invention relates tothe antibodies described herein, comprising the sequences of the V_(H),V_(L) and CDR polypeptides described herein, and the polynucleotidesencoding them. A preferred embodiment of the invention is directed tochimeric or humanized antibodies and fragments thereof (including Fabfragments) capable of binding to CGRP and/or inhibiting the biologicalactivities mediated by the binding of CGRP to the CGRP receptor(“CGRP-R”).

In another preferred embodiment of the invention, full length antibodiesand Fab fragments thereof are contemplated that inhibit the CGRP-alpha-,CGRP-beta-, and rat CGRP-driven production of cAMP. In a furtherpreferred embodiment of the invention, full length and Fab fragmentsthereof are contemplated that reduce vasodilation in a recipientfollowing administration.

In another embodiment of the invention, chimeric or humanized antibodiesand fragments thereof (including Fab fragments) capable of binding toCGRP are useful in methods directed to reducing, treating, or preventingmigraines (with or without aura), cancer or tumors, angiogenesisassociated with cancer or tumor growth, angiogenesis associated withcancer or tumor survival, weight loss, pain, hemiplagic migraines,cluster headaches, migrainous neuralgia, chronic headaches, tensionheadaches, general headaches, hot flushes, chronic paroxysomalhemicrania, secondary headaches due to an underlying structural problemin the head or neck, cranial neuralgia, sinus headaches (such as forexample associated with sinusitis), and allergy-induced headaches ormigraines. The antibodies and antibody fragments of the presentinvention particularly have utility in treating, preventing,ameliorating, controlling or reducing the risk of one or more of thefollowing conditions or diseases: overactive bladder and other urinaryconditions including bladder infection, pain; chronic pain; neurogenicinflammation and inflammatory pain; neuropathic pain; eye pain; toothpain; post-surgical pain, trauma related pain, burn related pain,diabetes; non-insulin dependent diabetes mellitus and other inflammatoryautoimmune disorders, vascular disorders; inflammation; arthritis;bronchial hyperreactivity, asthma; shock; sepsis; opiate withdrawalsyndrome; morphine tolerance; hot flashes in men and women; allergicdermatitis; psoriasis; encephalitis; brain trauma; epilepsy;neurodegenerative diseases; skin diseases including pruritis, neurogeniccutaneous redness, skin rosaceousness and erythema; inflammatory boweldisease, irritable bowel syndrome, cystitis; dysmenorrhea, and otherconditions that potentially may be treated or prevented or the symptomsameliorated by antagonism of CGRP receptors. Of particular importance isthe acute or prophylactic treatment of headache, including migraine andcluster headache, and other pain related conditions as well asoveractive bladder.

In another embodiment of the invention, chimeric or humanized antibodiesand fragments thereof (including Fab fragments) capable of binding toCGRP are preferably useful in methods directed to reducing, treating, orpreventing gastro-esophageal reflux, and visceral pain associated withgastro-esophageal reflux, dyspepsia, irritable bowel syndrome,inflammatory bowel disease, Crohn's disease, ileitis, ulcerativecolitis, renal colic, dysmenorrhea, cystitis, menstrual period, labor,menopause, prostatitis, or pancreatitis.

In another embodiment of the invention these antibodies and humanizedversions may be derived from rabbit immune cells (B lymphocytes) and maybe selected based on their homology (sequence identity) to human germline sequences. These antibodies may require minimal or no sequencemodifications, thereby facilitating retention of functional propertiesafter humanization. A further embodiment of the invention is directed tofragments from anti-CGRP antibodies encompassing V_(H), V_(L) and CDRpolypeptides, e.g., derived from rabbit immune cells and thepolynucleotides encoding the same, as well as the use of these antibodyfragments and the polynucleotides encoding them in the creation of novelantibodies and polypeptide compositions capable of binding to CGRPand/or CGRP/CGRP-R complexes.

The invention also contemplates conjugates of anti-CGRP antibodies andbinding fragments thereof conjugated to one or more functional ordetectable moieties. The invention also contemplates methods of makingsaid chimeric or humanized anti-CGRP or anti-CGRP/CGRP-R complexantibodies and binding fragments thereof. In one embodiment, bindingfragments include, but are not limited to, Fab, Fab′, F(ab′)₂, Fv, scFvfragments, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, and IgNAR.

Embodiments of the invention pertain to the use of anti-CGRP antibodiesand binding fragments thereof for the diagnosis, assessment andtreatment of diseases and disorders associated with CGRP or aberrantexpression thereof. The invention also contemplates the use of fragmentsof anti-CGRP antibodies for the diagnosis, assessment and treatment ofdiseases and disorders associated with CGRP or aberrant expressionthereof. Other embodiments of the invention relate to the production ofanti-CGRP antibodies or fragments thereof in recombinant host cells, forexample mammalian cells such as CHO, NSO or HEK 293 cells, or yeastcells (for example diploid yeast such as diploid Pichia) and other yeaststrains.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

FIG. 1 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab1.

FIG. 2 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab2.

FIG. 3 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab3.

FIG. 4 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab4.

FIG. 5 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab5.

FIG. 6 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab6.

FIG. 7 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab7.

FIG. 8 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab8.

FIG. 9 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab9.

FIG. 10 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab10.

FIG. 11 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab11.

FIG. 12 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab12.

FIG. 13 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab13.

FIG. 14 provides polynucleotide and polypeptide sequences correspondingto the full-length Antibody Ab14.

FIG. 15 provides the CGRP-alpha ELISA binding data obtained followingthe protocol in Example 1 infra for antibodies Ab1, Ab2, Ab3, and Ab4.

FIG. 16 provides the CGRP-alpha ELISA binding data obtained followingthe protocol in Example 1 infra for antibodies Ab5, Ab6, Ab7, and Ab8.

FIG. 17 provides the CGRP-alpha ELISA binding data obtained followingthe protocol in Example 1 infra for antibodies Ab9, Ab10, and Ab14.

FIG. 18 provides the CGRP-alpha ELISA binding data obtained followingthe protocol in Example 1 infra for antibodies Ab11, Ab12, and Ab13.

FIG. 19 demonstrates the inhibition of CGRP-alpha-driven cAMP productionby antibodies Ab1, Ab2, and Ab4, obtained following the protocol inExample 1 infra.

FIG. 20 demonstrates the inhibition of CGRP-alpha-driven cAMP productionby antibody Ab3, obtained following the protocol in Example 1 infra.

FIG. 21 demonstrates the inhibition of CGRP-alpha-driven cAMP productionby antibodies Ab5 and Ab6, obtained following the protocol in Example 1infra.

FIG. 22 demonstrates the inhibition of CGRP-alpha-driven cAMP productionby antibodies Ab7, Ab8, Ab9, and Ab10, obtained following the protocolin Example 1 infra.

FIG. 23 demonstrates the inhibition of CGRP-alpha-driven cAMP productionby antibodies Ab11, Ab12, and Ab13, obtained following the protocol inExample 1 infra.

FIG. 24 demonstrates the inhibition of CGRP-alpha-driven cAMP productionby antibody Ab14, obtained following the protocol in Example 1 infra.

FIG. 25 demonstrates the inhibition of CGRP-beta-driven cAMP productionby antibodies Ab1, Ab2, and Ab3, obtained following the protocol inExample 1 infra.

FIG. 26 demonstrates the inhibition of CGRP-beta-driven cAMP productionby antibodies Ab4, Ab5, and Ab6, obtained following the protocol inExample 1 infra.

FIG. 27 demonstrates the inhibition of CGRP-beta-driven cAMP productionby antibodies Ab7 and Ab8, obtained following the protocol in Example 1infra.

FIG. 28 demonstrates the inhibition of CGRP-beta-driven cAMP productionby antibodies Ab9, Ab10, and Ab14, obtained following the protocol inExample 1 infra.

FIG. 29 demonstrates the inhibition of CGRP-beta-driven cAMP productionby antibodies Ab11, Ab12, and Ab13, obtained following the protocol inExample 1 infra.

FIG. 30 demonstrates the inhibition of rat CGRP-driven cAMP productionby antibodies Ab1, Ab2, Ab4, and Ab5, obtained following the protocol inExample 1 infra.

FIG. 31 demonstrates the inhibition of rat CGRP-driven cAMP productionby antibodies Ab3 and Ab6, obtained following the protocol in Example 1infra.

FIG. 32 demonstrates the inhibition of rat CGRP-driven cAMP productionby antibodies Ab7 and Ab8, obtained following the protocol in Example 1infra.

FIG. 33 demonstrates the inhibition of rat CGRP-driven cAMP productionby antibody Ab9, obtained following the protocol in Example 1 infra.

FIG. 34 demonstrates the inhibition of rat CGRP-driven cAMP productionby antibody Ab10, obtained following the protocol in Example 1 infra.

FIG. 35 demonstrates the inhibition of rat CGRP-driven cAMP productionby antibodies Ab11 and Ab12, obtained following the protocol in Example1 infra.

FIG. 36 demonstrates the inhibition of rat CGRP-driven cAMP productionby antibody Ab13, obtained following the protocol in Example 1 infra.

FIG. 37 demonstrates the inhibition of rat CGRP-driven cAMP productionby antibody Ab14, obtained following the protocol in Example 1 infra.

FIG. 38 demonstrates the inhibition of binding of radiolabeled CGRP toCGRP-R by antibodies Ab1-Ab13, obtained following the protocol inExample 6 infra.

FIG. 39 demonstrates a reduction in vasodilation obtained byadministering antibodies Ab3 and Ab6 following capsaicin administrationin a rat model, relative to a control antibody, obtained following theprotocol in Example 7 infra.

FIG. 40 demonstrates a reduction in vasodilation obtained byadministering antibody Ab6 at differing concentrations followingcapsaicin administration in a rat model, relative to a control antibody,obtained following the protocol in Example 7 infra.

FIG. 41A-C shows the beneficial effect of Ab3 on bladder capacity duringsaline infusion. Animals were administered Ab3 or a negative controlantibody, then monitored during infusion of saline into the bladder. ICI(panel A) was increased and MF (panel B) was decreased, indicatingincreased bladder capacity. Differences in AM (panel C) were within thestandard deviation and not statistically significant. Asterisks indicatestatistically significant improvement (p<0.05 unpaired Student's t-test,comparison to Negative control Ab). Legend: filled bars: Ab3 treated (10mg/kg); open bars: negative control antibody (10 mg/kg). Error barsindicate the standard deviation. Abbreviations: ICI: IntercontractionInterval; MF: Micturition Frequency; AM: Amplitude of Micturition.

FIG. 42 shows the effect Ab2 in a model of neuropathic pain. Mechanicalallodynia was induced by Chung surgery (L5/L6 spinal nerve ligation),and degree of sensitivity was compared between Ab2 treated animals(hashed bars) and control animals (filled bars). Higher values indicateless sensitivity. Average sensitivity was similar at day 13 (prior toAb2 administration) but improved at days 14 and 17. Error bars indicatethe standard error of the mean.

FIG. 43 shows the analgesic effect of Ab2 and morphine. Pain sensitivitywas assessed by the tail withdrawal time (y-axis, seconds) for animalsadministered morphine (open squares), Ab2 (10 mg/kg, filled triangles),or vehicle (negative control, open circles). Animals developed morphinetolerance and exhibited tail withdrawal times similar to control animalsby day 4. In contrast, Ab2-treated animals exhibited a sustainedimprovement in tail withdrawal time over the course of the experiment(to day 7). The improvement in Ab2-treated animals was statisticallysignificant (p<0.05 one-way ANOVA followed by Dunnett's test, comparisonto vehicle, indicated by asterisks). Error bars indicate the standarderror of the mean.

FIG. 44 shows the dosage-dependent analgesic effect of Ab2. On day 0(subsequent to the first tail withdrawal time test), rats wereadministered antibody Ab2 at a dosage of 1 mg/kg (filled squares), 3mg/kg (filled downward-pointing triangles), or 10 mg/kg (filledupward-pointing triangles), or a vehicle (open circles) or negativecontrol antibody (open squares). The rats' tail withdrawal time inresponse to a painful thermal stimulus was assessed daily (higher timesindicate relative insensitivity to pain). Tail withdrawal time wasincreased in a dosage-dependent manner by the administration of Ab2.Asterisks indicate statistically significant increases in tailwithdrawal time (p<0.05 one-way ANOVA followed by Dunnett's test,comparison to vehicle). Error bars indicate the standard error of themean.

FIG. 45 shows the analgesic effect of Ab2 in combination with morphine,and when morphine is withdrawn after the onset of morphine tolerance. Onday 0 (subsequent to the first tail withdrawal time test), rats wereadministered antibody Ab2 at a dosage of 10 mg/kg (filled squares andfilled triangles) or vehicle (open circles). The rats were then dailyadministered morphine on days 1 to 10 (filled squares) or only on days 1to 4 (filled triangles). The tail withdrawal time initially was greatlyincreased in the morphine-treated mice, but decreased on subsequent daysindicating morphine tolerance. However, in the mice from which morphinewas withdrawn after day 4, tail withdrawal time increased and remainedelevated between days 5 and 8. Error bars indicate the standard error ofthe mean.

FIG. 46 shows the effect of Ab2 in a rat model of visceral pain.Visceral pain was quantified by measuring the colonic distensionthreshold (higher values indicate less sensitivity) for naïve animals(open bar) or animals treated with TNBS to provoke chronic colonichypersensitivity which either received a negative control antibody(filled bars) or Ab2 (hashed bars). Hypersensitivity was alleviated by27% by the Ab2-treated animals, and distension threshold wassignificantly improved by administration of Ab2 (p<0.05 Student'st-test, comparison to TNBS+Negative control group). Error bars indicatethe standard error of the mean.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS Definitions

It is to be understood that this invention is not limited to theparticular methodology, protocols, cell lines, animal species or genera,and reagents described, as such may vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to limit the scope ofthe present invention which will be limited only by the appended claims.As used herein the singular forms “a”, “and”, and “the” include pluralreferents unless the context clearly dictates otherwise. Thus, forexample, reference to “a cell” includes a plurality of such cells andreference to “the protein” includes reference to one or more proteinsand equivalents thereof known to those skilled in the art, and so forth.All technical and scientific terms used herein have the same meaning ascommonly understood to one of ordinary skill in the art to which thisinvention belongs unless clearly indicated otherwise.

Calcitonin Gene Related Peptide (CGRP): As used herein, CGRP encompassesnot only the following Homo sapiens CGRP-alpha and Homo sapiensCGRP-beta amino acid sequences available from American Peptides(Sunnyvale Calif.) and Bachem (Torrance, Calif.): CGRP-alpha:ACDTATCVTHRLAGLLSRSGGVVKNNFVPTNVGSKAF-NH₂ (SEQ ID NO: 281), wherein theN-terminal phenylalanine is amidated;

CGRP-beta: ACNTATCVTHRLAGLLSRSGGMVKSNFVPTNVGSKAF-NH₂ (SEQ ID NO: 282),wherein the N-terminal phenylalanine is amidated; but also anymembrane-bound forms of these CGRP amino acid sequences, as well asmutants (mutiens), splice variants, isoforms, orthologues, homologuesand variants of this sequence.

Mating competent yeast species: In the present invention this isintended to broadly encompass any diploid or tetraploid yeast which canbe grown in culture. Such species of yeast may exist in a haploid,diploid, or other polyploid form. The cells of a given ploidy may, underappropriate conditions, proliferate for an indefinite number ofgenerations in that form. Diploid cells can also sporulate to formhaploid cells. Sequential mating can result in tetraploid strainsthrough further mating or fusion of diploid strains. The presentinvention contemplates the use of haploid yeast, as well as diploid orother polyploid yeast cells produced, for example, by mating orspheroplast fusion.

In one embodiment of the invention, the mating competent yeast is amember of the Saccharomycetaceae family, which includes the generaArxiozyma; Ascobotryozyma; Citeromyces; Debaryomyces; Dekkera;Eremothecium; Issatchenkia; Kazachstania; Kluyveromyces; Kodamaea;Lodderomyces; Pachysolen; Pichia; Saccharomyces; Saturnispora;Tetrapisispora; Torulaspora; Williopsis; and Zygosaccharomyces. Othertypes of yeast potentially useful in the invention include Yarrowia;Rhodosporidium; Candida; Hansenula; Filobasium; Sporidiobolus; Bullera;Leucosporidium and Filobasidella.

In a preferred embodiment of the invention, the mating competent yeastis a member of the genus Pichia. In a further preferred embodiment ofthe invention, the mating competent yeast of the genus Pichia is one ofthe following species: Pichia pastoris, Pichia methanolica, andHansenula polymorpha (Pichia angusta). In a particularly preferredembodiment of the invention, the mating competent yeast of the genusPichia is the species Pichia pastoris.

Haploid Yeast Cell: A cell having a single copy of each gene of itsnormal genomic (chromosomal) complement.

Polyploid Yeast Cell: A cell having more than one copy of its normalgenomic (chromosomal) complement.

Diploid Yeast Cell: A cell having two copies (alleles) of essentiallyevery gene of its normal genomic complement, typically formed by theprocess of fusion (mating) of two haploid cells.

Tetraploid Yeast Cell: A cell having four copies (alleles) ofessentially every gene of its normal genomic complement, typicallyformed by the process of fusion (mating) of two haploid cells.Tetraploids may carry two, three, four or more different expressioncassettes. Such tetraploids might be obtained in S. cerevisiae byselective mating homozygotic heterothallic a/a and alpha/alpha diploidsand in Pichia by sequential mating of haploids to obtain auxotrophicdiploids. For example, a [met his] haploid can be mated with [ade his]haploid to obtain diploid [his]; and a [met arg] haploid can be matedwith [ade arg] haploid to obtain diploid [arg]; then the diploid [his] xdiploid [arg] to obtain a tetraploid prototroph. It will be understoodby those of skill in the art that reference to the benefits and uses ofdiploid cells may also apply to tetraploid cells.

Yeast Mating: The process by which two haploid yeast cells naturallyfuse to form one diploid yeast cell.

Meiosis: The process by which a diploid yeast cell undergoes reductivedivision to form four haploid spore products. Each spore may thengerminate and form a haploid vegetatively growing cell line.

Selectable Marker: A selectable marker is a gene or gene fragment thatconfers a growth phenotype (physical growth characteristic) on a cellreceiving that gene as, for example through a transformation event. Theselectable marker allows that cell to survive and grow in a selectivegrowth medium under conditions in which cells that do not receive thatselectable marker gene cannot grow. Selectable marker genes generallyfall into several types, including positive selectable marker genes suchas a gene that confers on a cell resistance to an antibiotic or otherdrug, temperature when two temperature sensitive (“ts”) mutants arecrossed or a is mutant is transformed; negative selectable marker genessuch as a biosynthetic gene that confers on a cell the ability to growin a medium without a specific nutrient needed by all cells that do nothave that biosynthetic gene, or a mutagenized biosynthetic gene thatconfers on a cell inability to grow by cells that do not have the wildtype gene; and the like. Suitable markers include but are not limitedto: ZEO; G418; LYS3; MET1; MET3a; ADE1; ADE3; URA3; and the like.

Expression Vector: These DNA vectors contain elements that facilitatemanipulation for the expression of a foreign protein within the targethost cell. Conveniently, manipulation of sequences and production of DNAfor transformation is first performed in a bacterial host, e.g. E. coli,and usually vectors will include sequences to facilitate suchmanipulations, including a bacterial origin of replication andappropriate bacterial selection marker. Selection markers encodeproteins necessary for the survival or growth of transformed host cellsgrown in a selective culture medium. Host cells not transformed with thevector containing the selection gene will not survive in the culturemedium. Typical selection genes encode proteins that (a) conferresistance to antibiotics or other toxins, (b) complement auxotrophicdeficiencies, or (c) supply critical nutrients not available fromcomplex media. Exemplary vectors and methods for transformation of yeastare described, for example, in Burke, D., Dawson, D., & Stearns, T.(2000). Methods in yeast genetics: a Cold Spring Harbor Laboratorycourse manual. Plainview, N.Y.: Cold Spring Harbor Laboratory Press.

Expression vectors for use in the methods of the invention will furtherinclude yeast specific sequences, including a selectable auxotrophic ordrug marker for identifying transformed yeast strains. A drug marker mayfurther be used to amplify copy number of the vector in a yeast hostcell.

The polypeptide coding sequence of interest is operably linked totranscriptional and translational regulatory sequences that provide forexpression of the polypeptide in yeast cells. These vector componentsmay include, but are not limited to, one or more of the following: anenhancer element, a promoter, and a transcription termination sequence.Sequences for the secretion of the polypeptide may also be included,e.g. a signal sequence, and the like. A yeast origin of replication isoptional, as expression vectors are often integrated into the yeastgenome. In one embodiment of the invention, the polypeptide of interestis operably linked, or fused, to sequences providing for optimizedsecretion of the polypeptide from yeast diploid cells.

Nucleic acids are “operably linked” when placed into a functionalrelationship with another nucleic acid sequence. For example, DNA for asignal sequence is operably linked to DNA for a polypeptide if it isexpressed as a preprotein that participates in the secretion of thepolypeptide; a promoter or enhancer is operably linked to a codingsequence if it affects the transcription of the sequence. Generally,“operably linked” means that the DNA sequences being linked arecontiguous, and, in the case of a secretory leader, contiguous and inreading frame. However, enhancers do not have to be contiguous. Linkingis accomplished by ligation at convenient restriction sites oralternatively via a PCR/recombination method familiar to those skilledin the art (Gateway® Technology; Invitrogen, Carlsbad Calif.). If suchsites do not exist, the synthetic oligonucleotide adapters or linkersare used in accordance with conventional practice.

Promoters are untranslated sequences located upstream (5′) to the startcodon of a structural gene (generally within about 100 to 1000 bp) thatcontrol the transcription and translation of particular nucleic acidsequences to which they are operably linked. Such promoters fall intoseveral classes: inducible, constitutive, and repressible promoters(that increase levels of transcription in response to absence of arepressor). Inducible promoters may initiate increased levels oftranscription from DNA under their control in response to some change inculture conditions, e.g., the presence or absence of a nutrient or achange in temperature.

The yeast promoter fragment may also serve as the site for homologousrecombination and integration of the expression vector into the samesite in the yeast genome; alternatively a selectable marker is used asthe site for homologous recombination. Pichia transformation isdescribed in Cregg et al. (1985) Mol. Cell. Biol. 5:3376-3385.

Examples of suitable promoters from Pichia include the AOX1 and promoter(Cregg et al. (1989) Mol. Cell. Biol. 9:1316-1323); ICL1 promoter(Menendez et al. (2003) Yeast 20(13):1097-108);glyceraldehyde-3-phosphate dehydrogenase promoter (GAP) (Waterham et al.(1997) Gene 186(1):37-44); and FLD1 promoter (Shen et al. (1998) Gene216(1):93-102). The GAP promoter is a strong constitutive promoter andthe AOX and FLD1 promoters are inducible.

Other yeast promoters include ADH1, alcohol dehydrogenase II, GAL4,PHO3, PHO5, Pyk, and chimeric promoters derived therefrom. Additionally,non-yeast promoters may be used in the invention such as mammalian,insect, plant, reptile, amphibian, viral, and avian promoters. Mosttypically the promoter will comprise a mammalian promoter (potentiallyendogenous to the expressed genes) or will comprise a yeast or viralpromoter that provides for efficient transcription in yeast systems.

The polypeptides of interest may be produced recombinantly not onlydirectly, but also as a fusion polypeptide with a heterologouspolypeptide, e.g. a signal sequence or other polypeptide having aspecific cleavage site at the N-terminus of the mature protein orpolypeptide. In general, the signal sequence may be a component of thevector, or it may be a part of the polypeptide coding sequence that isinserted into the vector. The heterologous signal sequence selectedpreferably is one that is recognized and processed through one of thestandard pathways available within the host cell. The S. cerevisiaealpha factor pre-pro signal has proven effective in the secretion of avariety of recombinant proteins from P. pastoris. Other yeast signalsequences include the alpha mating factor signal sequence, the invertasesignal sequence, and signal sequences derived from other secreted yeastpolypeptides. Additionally, these signal peptide sequences may beengineered to provide for enhanced secretion in diploid yeast expressionsystems. Other secretion signals of interest also include mammaliansignal sequences, which may be heterologous to the protein beingsecreted, or may be a native sequence for the protein being secreted.Signal sequences include pre-peptide sequences, and in some instancesmay include propeptide sequences. Many such signal sequences are knownin the art, including the signal sequences found on immunoglobulinchains, e.g., K28 preprotoxin sequence, PHA-E, FACE, human MCP-1, humanserum albumin signal sequences, human Ig heavy chain, human Ig lightchain, and the like. For example, see Hashimoto et. al. Protein Eng11(2) 75 (1998); and Kobayashi et. al. Therapeutic Apheresis 2(4) 257(1998).

Transcription may be increased by inserting a transcriptional activatorsequence into the vector. These activators are cis-acting elements ofDNA, usually about from 10 to 300 bp, which act on a promoter toincrease its transcription. Transcriptional enhancers are relativelyorientation and position independent, having been found 5′ and 3′ to thetranscription unit, within an intron, as well as within the codingsequence itself. The enhancer may be spliced into the expression vectorat a position 5′ or 3′ to the coding sequence, but is preferably locatedat a site 5′ from the promoter. Expression vectors used in eukaryotichost cells may also contain sequences necessary for the termination oftranscription and for stabilizing the mRNA. Such sequences are commonlyavailable from 3′ to the translation termination codon, in untranslatedregions of eukaryotic or viral DNAs or cDNAs. These regions containnucleotide segments transcribed as polyadenylated fragments in theuntranslated portion of the mRNA.

Construction of suitable vectors containing one or more of theabove-listed components employs standard ligation techniques orPCR/recombination methods. Isolated plasmids or DNA fragments arecleaved, tailored, and re-ligated in the form desired to generate theplasmids required or via recombination methods. For analysis to confirmcorrect sequences in plasmids constructed, the ligation mixtures areused to transform host cells, and successful transformants selected byantibiotic resistance (e.g. ampicillin or Zeocin) where appropriate.Plasmids from the transformants are prepared, analyzed by restrictionendonuclease digestion and/or sequenced.

As an alternative to restriction and ligation of fragments,recombination methods based on att sites and recombination enzymes maybe used to insert DNA sequences into a vector. Such methods aredescribed, for example, by Landy (1989) Ann. Rev. Biochem. 58:913-949;and are known to those of skill in the art. Such methods utilizeintermolecular DNA recombination that is mediated by a mixture of lambdaand E. coli—encoded recombination proteins. Recombination occurs betweenspecific attachment (att) sites on the interacting DNA molecules. For adescription of att sites see Weisberg and Landy (1983) Site-SpecificRecombination in Phage Lambda, in Lambda II, Weisberg, ed. (Cold SpringHarbor, N.Y.: Cold Spring Harbor Press), pp. 211-250. The DNA segmentsflanking the recombination sites are switched, such that afterrecombination, the att sites are hybrid sequences comprised of sequencesdonated by each parental vector. The recombination can occur betweenDNAs of any topology.

Att sites may be introduced into a sequence of interest by ligating thesequence of interest into an appropriate vector; generating a PCRproduct containing att B sites through the use of specific primers;generating a cDNA library cloned into an appropriate vector containingatt sites; and the like.

Folding, as used herein, refers to the three-dimensional structure ofpolypeptides and proteins, where interactions between amino acidresidues act to stabilize the structure. While non-covalent interactionsare important in determining structure, usually the proteins of interestwill have intra- and/or intermolecular covalent disulfide bonds formedby two cysteine residues. For naturally occurring proteins andpolypeptides or derivatives and variants thereof, the proper folding istypically the arrangement that results in optimal biological activity,and can conveniently be monitored by assays for activity, e.g. ligandbinding, enzymatic activity, etc.

In some instances, for example where the desired product is of syntheticorigin, assays based on biological activity will be less meaningful. Theproper folding of such molecules may be determined on the basis ofphysical properties, energetic considerations, modeling studies, and thelike.

The expression host may be further modified by the introduction ofsequences encoding one or more enzymes that enhance folding anddisulfide bond formation, i.e. foldases, chaperonins, etc. Suchsequences may be constitutively or inducibly expressed in the yeast hostcell, using vectors, markers, etc. as known in the art. Preferably thesequences, including transcriptional regulatory elements sufficient forthe desired pattern of expression, are stably integrated in the yeastgenome through a targeted methodology.

For example, the eukaryotic PDI is not only an efficient catalyst ofprotein cysteine oxidation and disulfide bond isomerization, but alsoexhibits chaperone activity. Co-expression of PDI can facilitate theproduction of active proteins having multiple disulfide bonds. Also ofinterest is the expression of BIP (immunoglobulin heavy chain bindingprotein); cyclophilin; and the like. In one embodiment of the invention,each of the haploid parental strains expresses a distinct foldingenzyme, e.g. one strain may express BIP, and the other strain mayexpress PDI or combinations thereof.

The terms “desired protein” or “desired antibody” are usedinterchangeably and refer generally to a parent antibody specific to atarget, i.e., CGRP or a chimeric or humanized antibody or a bindingportion thereof derived therefrom as described herein. The term“antibody” is intended to include any polypeptide chain-containingmolecular structure with a specific shape that fits to and recognizes anepitope, where one or more non-covalent binding interactions stabilizethe complex between the molecular structure and the epitope. Thearchetypal antibody molecule is the immunoglobulin, and all types ofimmunoglobulins, IgG, IgM, IgA, IgE, IgD, etc., from all sources, e.g.human, rodent, rabbit, cow, sheep, pig, dog, other mammals, chicken,other avians, etc., are considered to be “antibodies.” A preferredsource for producing antibodies useful as starting material according tothe invention is rabbits. Numerous antibody coding sequences have beendescribed; and others may be raised by methods well-known in the art.Examples thereof include chimeric antibodies, human antibodies and othernon-human mammalian antibodies, humanized antibodies, single chainantibodies (such as scFvs), camelbodies, nanobodies, IgNAR (single-chainantibodies derived from sharks), small-modular immunopharmaceuticals(SMIPs), and antibody fragments such as Fabs, Fab′, F(ab′)₂ and thelike. See Streltsov V A, et al., Structure of a shark IgNAR antibodyvariable domain and modeling of an early-developmental isotype, ProteinSci. 2005 November; 14(11):2901-9. Epub 2005 Sep 30; Greenberg AS, etal., A new antigen receptor gene family that undergoes rearrangement andextensive somatic diversification in sharks, Nature. 1995 Mar. 9;374(6518):168-73; Nuttall S D, et al., Isolation of the new antigenreceptor from wobbegong sharks, and use as a scaffold for the display ofprotein loop libraries, Mol. Immunol. 2001 August; 38(4):313-26;Hamers-Casterman C, et al., Naturally occurring antibodies devoid oflight chains, Nature. 1993 Jun. 3; 363(6428):446-8; Gill D S, et al.,Biopharmaceutical drug discovery using novel protein scaffolds, CurrOpin Biotechnol. December; 17(6):653-8. Epub 2006 Oct 19.

For example, antibodies or antigen binding fragments may be produced bygenetic engineering. In this technique, as with other methods,antibody-producing cells are sensitized to the desired antigen orimmunogen. The messenger RNA isolated from antibody producing cells isused as a template to make cDNA using PCR amplification. A library ofvectors, each containing one heavy chain gene and one light chain generetaining the initial antigen specificity, is produced by insertion ofappropriate sections of the amplified immunoglobulin cDNA into theexpression vectors. A combinatorial library is constructed by combiningthe heavy chain gene library with the light chain gene library. Thisresults in a library of clones which co-express a heavy and light chain(resembling the Fab fragment or antigen binding fragment of an antibodymolecule). The vectors that carry these genes are co-transfected into ahost cell. When antibody gene synthesis is induced in the transfectedhost, the heavy and light chain proteins self-assemble to produce activeantibodies that can be detected by screening with the antigen orimmunogen.

Antibody coding sequences of interest include those encoded by nativesequences, as well as nucleic acids that, by virtue of the degeneracy ofthe genetic code, are not identical in sequence to the disclosed nucleicacids, and variants thereof. Variant polypeptides can include amino acid(aa) substitutions, additions or deletions. The amino acid substitutionscan be conservative amino acid substitutions or substitutions toeliminate non-essential amino acids, such as to alter a glycosylationsite, or to minimize misfolding by substitution or deletion of one ormore cysteine residues that are not necessary for function. Variants canbe designed so as to retain or have enhanced biological activity of aparticular region of the protein (e.g., a functional domain, catalyticamino acid residues, etc). Variants also include fragments of thepolypeptides disclosed herein, particularly biologically activefragments and/or fragments corresponding to functional domains.Techniques for in vitro mutagenesis of cloned genes are known. Alsoincluded in the subject invention are polypeptides that have beenmodified using ordinary molecular biological techniques so as to improvetheir resistance to proteolytic degradation or to optimize solubilityproperties or to render them more suitable as a therapeutic agent.

Chimeric antibodies may be made by recombinant means by combining thevariable light and heavy chain regions (V_(L) and V_(H)), obtained fromantibody producing cells of one species with the constant light andheavy chain regions from another. Typically chimeric antibodies utilizerodent or rabbit variable regions and human constant regions, in orderto produce an antibody with predominantly human domains. The productionof such chimeric antibodies is well known in the art, and may beachieved by standard means (as described, e.g., in U.S. Pat. No.5,624,659, incorporated herein by reference in its entirety). It isfurther contemplated that the human constant regions of chimericantibodies of the invention may be selected from IgG1, IgG2, IgG3, andIgG4 constant regions.

Humanized antibodies are engineered to contain even more human-likeimmunoglobulin domains, and incorporate only thecomplementarity-determining regions of the animal-derived antibody. Thisis accomplished by carefully examining the sequence of thehyper-variable loops of the variable regions of the monoclonal antibody,and fitting them to the structure of the human antibody chains. Althoughfacially complex, the process is straightforward in practice. See, e.g.,U.S. Pat. No. 6,187,287, incorporated fully herein by reference.

In addition to entire immunoglobulins (or their recombinantcounterparts), immunoglobulin fragments comprising the epitope bindingsite (e.g., Fab′, F(ab′)₂, or other fragments) may be synthesized.“Fragment,” or minimal immunoglobulins may be designed utilizingrecombinant immunoglobulin techniques. For instance “Fv” immunoglobulinsfor use in the present invention may be produced by synthesizing a fusedvariable light chain region and a variable heavy chain region.Combinations of antibodies are also of interest, e.g. diabodies, whichcomprise two distinct Fv specificities. In another embodiment of theinvention, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, and IgNAR are encompassed by immunoglobulin fragments.

Immunoglobulins and fragments thereof may be modifiedpost-translationally, e.g. to add effector moieties such as chemicallinkers, detectable moieties, such as fluorescent dyes, enzymes, toxins,substrates, bioluminescent materials, radioactive materials,chemiluminescent moieties and the like, or specific binding moieties,such as streptavidin, avidin, or biotin, and the like may be utilized inthe methods and compositions of the present invention. Examples ofadditional effector molecules are provided infra.

A polynucleotide sequence “corresponds” to a polypeptide sequence iftranslation of the polynucleotide sequence in accordance with thegenetic code yields the polypeptide sequence (i.e., the polynucleotidesequence “encodes” the polypeptide sequence), one polynucleotidesequence “corresponds” to another polynucleotide sequence if the twosequences encode the same polypeptide sequence.

A “heterologous” region or domain of a DNA construct is an identifiablesegment of DNA within a larger DNA molecule that is not found inassociation with the larger molecule in nature. Thus, when theheterologous region encodes a mammalian gene, the gene will usually beflanked by DNA that does not flank the mammalian genomic DNA in thegenome of the source organism. Another example of a heterologous regionis a construct where the coding sequence itself is not found in nature(e.g., a cDNA where the genomic coding sequence contains introns, orsynthetic sequences having codons different than the native gene).Allelic variations or naturally-occurring mutational events do not giverise to a heterologous region of DNA as defined herein.

A “coding sequence” is an in-frame sequence of codons that (in view ofthe genetic code) correspond to or encode a protein or peptide sequence.Two coding sequences correspond to each other if the sequences or theircomplementary sequences encode the same amino acid sequences. A codingsequence in association with appropriate regulatory sequences may betranscribed and translated into a polypeptide. A polyadenylation signaland transcription termination sequence will usually be located 3′ to thecoding sequence. A “promoter sequence” is a DNA regulatory regioncapable of binding RNA polymerase in a cell and initiating transcriptionof a downstream (3′ direction) coding sequence. Promoter sequencestypically contain additional sites for binding of regulatory molecules(e.g., transcription factors) which affect the transcription of thecoding sequence. A coding sequence is “under the control” of thepromoter sequence or “operatively linked” to the promoter when RNApolymerase binds the promoter sequence in a cell and transcribes thecoding sequence into mRNA, which is then in turn translated into theprotein encoded by the coding sequence.

Vectors are used to introduce a foreign substance, such as DNA, RNA orprotein, into an organism or host cell. Typical vectors includerecombinant viruses (for polynucleotides) and liposomes (forpolypeptides). A “DNA vector” is a replicon, such as plasmid, phage orcosmid, to which another polynucleotide segment may be attached so as tobring about the replication of the attached segment. An “expressionvector” is a DNA vector which contains regulatory sequences which willdirect polypeptide synthesis by an appropriate host cell. This usuallymeans a promoter to bind RNA polymerase and initiate transcription ofmRNA, as well as ribosome binding sites and initiation signals to directtranslation of the mRNA into a polypeptide(s). Incorporation of apolynucleotide sequence into an expression vector at the proper site andin correct reading frame, followed by transformation of an appropriatehost cell by the vector, enables the production of a polypeptide encodedby said polynucleotide sequence.

“Amplification” of polynucleotide sequences is the in vitro productionof multiple copies of a particular nucleic acid sequence. The amplifiedsequence is usually in the form of DNA. A variety of techniques forcarrying out such amplification are described in a review article by VanBrunt (1990, Bio/Technol., 8(4):291-294). Polymerase chain reaction orPCR is a prototype of nucleic acid amplification, and use of PCR hereinshould be considered exemplary of other suitable amplificationtechniques.

The general structure of antibodies in vertebrates now is wellunderstood (Edelman, G. M., Ann. N.Y. Acad. Sci., 190: 5 (1971)).Antibodies consist of two identical light polypeptide chains ofmolecular weight approximately 23,000 daltons (the “light chain”), andtwo identical heavy chains of molecular weight 53,000-70,000 (the “heavychain”). The four chains are joined by disulfide bonds in a “Y”configuration wherein the light chains bracket the heavy chains startingat the mouth of the “Y” configuration. The “branch” portion of the “Y”configuration is designated the F_(ab) region; the stem portion of the“Y” configuration is designated the F_(c) region. The amino acidsequence orientation runs from the N-terminal end at the top of the “Y”configuration to the C-terminal end at the bottom of each chain. TheN-terminal end possesses the variable region having specificity for theantigen that elicited it, and is approximately 100 amino acids inlength, there being slight variations between light and heavy chain andfrom antibody to antibody.

The variable region is linked in each chain to a constant region thatextends the remaining length of the chain and that within a particularclass of antibody does not vary with the specificity of the antibody(i.e., the antigen eliciting it). There are five known major classes ofconstant regions that determine the class of the immunoglobulin molecule(IgG, IgM, IgA, IgD, and IgE corresponding to γ, μ, α, δ, and ε (gamma,mu, alpha, delta, or epsilon) heavy chain constant regions). Theconstant region or class determines subsequent effector function of theantibody, including activation of complement (Kabat, E. A., StructuralConcepts in Immunology and Immunochemistry, 2nd Ed., p. 413-436, Holt,Rinehart, Winston (1976)), and other cellular responses (Andrews, D. W.,et al., Clinical Immunobiology, pp 1-18, W. B. Sanders (1980); Kohl, S.,et al., Immunology, 48: 187 (1983)); while the variable regiondetermines the antigen with which it will react. Light chains areclassified as either κ (kappa) or λ (lambda). Each heavy chain class canbe prepared with either kappa or lambda light chain. The light and heavychains are covalently bonded to each other, and the “tail” portions ofthe two heavy chains are bonded to each other by covalent disulfidelinkages when the immunoglobulins are generated either by hybridomas orby B cells.

The expression “variable region” or “VR” refers to the domains withineach pair of light and heavy chains in an antibody that are involveddirectly in binding the antibody to the antigen. Each heavy chain has atone end a variable domain (V_(H)) followed by a number of constantdomains. Each light chain has a variable domain (V_(L)) at one end and aconstant domain at its other end; the constant domain of the light chainis aligned with the first constant domain of the heavy chain, and thelight chain variable domain is aligned with the variable domain of theheavy chain.

The expressions “complementarity determining region,” “hypervariableregion,” or “CDR” refer to one or more of the hyper-variable orcomplementarity determining regions (CDRs) found in the variable regionsof light or heavy chains of an antibody (See Kabat, E. A. et al.,Sequences of Proteins of Immunological Interest, National Institutes ofHealth, Bethesda, Md., (1987)). These expressions include thehypervariable regions as defined by Kabat et al. (“Sequences of Proteinsof Immunological Interest,” Kabat E., et al., US Dept. of Health andHuman Services, 1983) or the hypervariable loops in 3-dimensionalstructures of antibodies (Chothia and Lesk, J. Mol. Biol. 196 901-917(1987)). The CDRs in each chain are held in close proximity by frameworkregions and, with the CDRs from the other chain, contribute to theformation of the antigen binding site. Within the CDRs there are selectamino acids that have been described as the selectivity determiningregions (SDRs) which represent the critical contact residues used by theCDR in the antibody-antigen interaction (Kashmiri, S., Methods, 36:25-34(2005)).

The expressions “framework region” or “FR” refer to one or more of theframework regions within the variable regions of the light and heavychains of an antibody (See Kabat, E. A. et al., Sequences of Proteins ofImmunological Interest, National Institutes of Health, Bethesda, Md.,(1987)). These expressions include those amino acid sequence regionsinterposed between the CDRs within the variable regions of the light andheavy chains of an antibody.

Anti-CGRP Antibodies and Binding Fragments Thereof Having BindingActivity for CGRP Antibody Ab1

In one embodiment, the invention includes chimeric antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 1) QVLTQTASPVSAAVGSTVTINCQASQSVYDNNYLAWYQQKPGQPPKQLIYSTSTLASGVSSRFKGSGSGTQFTLTISDLECADAATYYCLGSYDCSSG DCFVFGGGTEVVVKR.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 2) QVLTQTASPVSAAVGSTVTINCQASQSVYDNNYLAWYQQKPGQPPKQLIYSTSTLASGVSSRFKGSGSGTQFTLTISDLECADAATYYCLGSYDCSSGDCFVFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 3) QSLEESGGRLVTPGTPLTLTCTVSGLDLSSYYMQWVRQAPGKGLEWIGVIGINDNTYYASWAKGRFTISRASSTTVDLKMTSLTTEDTATYFCARGDI WGPGTLVTVSS.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 4) QSLEESGGRLVTPGTPLTLTCTVSGLDLSSYYMQWVRQAPGKGLEWIGVIGINDNTYYASWAKGRFTISRASSTTVDLKMTSLTTEDTATYFCARGDIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVELEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKENWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFELYSKLTVDKSRWQQGNVESCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 5; SEQ ID NO: 6; and SEQ ID NO:7 which correspond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable light chain sequence of SEQ IDNO: 1 or the light chain sequence of SEQ ID NO: 2, and/or one or more ofthe polypeptide sequences of SEQ ID NO: 8; SEQ ID NO: 9; and SEQ ID NO:10 which correspond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable heavy chain sequence of SEQ IDNO: 3 or the heavy chain sequence of SEQ ID NO: 4, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention or fragments thereof comprise, oralternatively consist of, combinations of one or more of the CDRs, thevariable heavy and variable light chain sequences, and the heavy andlight chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 3 or SEQ ID NO: 4.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 5; SEQ ID NO:6; and SEQ ID NO: 7 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable light chainsequence of SEQ ID NO: 1 or the light chain sequence of SEQ ID NO: 2.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 8; SEQ ID NO:9; and SEQ ID NO: 10 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 3 or the heavy chain sequence of SEQ ID NO: 4.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 1; the variable heavy chain region of SEQ IDNO: 3; the complementarity-determining regions (SEQ ID NO: 5; SEQ ID NO:6; and SEQ ID NO: 7) of the variable light chain region of SEQ ID NO: 1;and the complementarity-determining regions (SEQ ID NO: 8; SEQ ID NO: 9;and SEQ ID NO: 10) of the variable heavy chain region of SEQ ID NO: 3.

In a particularly preferred embodiment of the invention, the chimericanti-CGRP antibody is Ab1, comprising, or alternatively consisting of,SEQ ID NO: 2 and SEQ ID NO: 4, and having at least one of the biologicalactivities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab1, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 1 and the variable heavy chain sequence ofSEQ ID NO: 3. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 1 and/or SEQ ID NO: 3in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab1.In another embodiment of the invention, anti-CGRP antibodies such as Ab1or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Ab2

In one embodiment, the invention includes humanized antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 11) QVLTQSPSSLSASVGDRVTINCQASQSVYDNNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSSG DCFVFGGGTKVEIKR.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 12) QVLTQSPSSLSASVGDRVTINCQASQSVYDNNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSSGDCFVFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

The invention further includes humanized antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 13) EVQLVESGGGLVQPGGSLRLSCAVSGLDLSSYYMQWVRQAPGKGLEWVGVIGINDNTYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARG DIWGQGTLVTVSS.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 14) EVQLVESGGGLVQPGGSLRLSCAVSGLDLSSYYMQWVRQAPGKGLEWVGVIGINDNTYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARGDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 15; SEQ ID NO: 16; and SEQ IDNO: 17 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 11 or the light chain sequence of SEQ ID NO: 12, and/or oneor more of the polypeptide sequences of SEQ ID NO: 18; SEQ ID NO: 19;and SEQ ID NO: 20 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 13 or the heavy chain sequence of SEQ ID NO: 14,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 11 or SEQ ID NO: 12. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 13 or SEQ ID NO: 14.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 15; SEQ IDNO: 16; and SEQ ID NO: 17 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 11 or the light chainsequence of SEQ ID NO: 12.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 18; SEQ IDNO: 19; and SEQ ID NO: 20 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 13 or the heavy chainsequence of SEQ ID NO: 14.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 11; the variable heavy chain region of SEQ IDNO: 13; the complementarity-determining regions (SEQ ID NO: 15; SEQ IDNO: 16; and SEQ ID NO: 17) of the variable light chain region of SEQ IDNO: 11; and the complementarity-determining regions (SEQ ID NO: 18; SEQID NO: 19; and SEQ ID NO: 20) of the variable heavy chain region of SEQID NO: 13.

In a particularly preferred embodiment of the invention, the humanizedanti-CGRP antibody is Ab2, comprising, or alternatively consisting of,SEQ ID NO: 12 and SEQ ID NO: 14, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab2, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 11 and the variable heavy chain sequence ofSEQ ID NO: 13. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 11 and/or SEQ ID NO: 13in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab2.In another embodiment of the invention, anti-CGRP antibodies such as Ab2or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Ab3

In one embodiment, the invention includes humanized antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 21) QVLTQSPSSLSASVGDRVTINCQASQSVYDNNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSSG DCFVFGGGTKVEIKR.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 22) QVLTQSPSSLSASVGDRVTINCQASQSVYDNNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSSGDCFVFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC.

The invention further includes humanized antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 23) EVQLVESGGGLVQPGGSLRLSCAVSGLDLSSYYMQWVRQAPGKGLEWVGVIGINDNTYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARGDI WGQGTLVTVSS.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 24) EVQLVESGGGLVQPGGSLRLSCAVSGLDLSSYYMQWVRQAPGKGLEWVGVIGINDNTYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARGDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 25; SEQ ID NO: 26; and SEQ IDNO: 27 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 21 or the light chain sequence of SEQ ID NO: 22, and/or oneor more of the polypeptide sequences of SEQ ID NO: 28; SEQ ID NO: 29;and SEQ ID NO: 30 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 23 or the heavy chain sequence of SEQ ID NO: 24,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 21 or SEQ ID NO: 22. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 23 or SEQ ID NO: 24.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 25; SEQ IDNO: 26; and SEQ ID NO: 27 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 21 or the light chainsequence of SEQ ID NO: 22.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 28; SEQ IDNO: 29; and SEQ ID NO: 30 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 23 or the heavy chainsequence of SEQ ID NO: 24.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 21; the variable heavy chain region of SEQ IDNO: 23; the complementarity-determining regions (SEQ ID NO: 25; SEQ IDNO: 26; and SEQ ID NO: 27) of the variable light chain region of SEQ IDNO: 21; and the complementarity-determining regions (SEQ ID NO: 28; SEQID NO: 29; and SEQ ID NO: 30) of the variable heavy chain region of SEQID NO: 23.

In a particularly preferred embodiment of the invention, the chimericanti-CGRP antibody is Ab3, comprising, or alternatively consisting of,SEQ ID NO: 22 and SEQ ID NO: 24, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab3, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 21 and the variable heavy chain sequence ofSEQ ID NO: 23. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 21 and/or SEQ ID NO: 23in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab3.In another embodiment of the invention, anti-CGRP antibodies such as Ab3or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Ab4

In one embodiment, the invention includes chimeric antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 31) QVLTQTPSPVSAAVGSTVTINCQASQSVYHNTYLAWYQQKPGQPPKQLIYDASTLASGVPSRFSGSGSGTQFTLTISGVQCNDAAAYYCLGSYDCTNGDC FVFGGGTEVVVKR.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 32) QVLTQTPSPVSAAVGSTVTINCQASQSVYHNTYLAWYQQKPGQPPKQLIYDASTLASGVPSRFSGSGSGTQFTLTISGVQCNDAAAYYCLGSYDCTNGDCFVFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 33) QSLEESGGRLVTPGTPLTLTCSVSGIDLSGYYMNWVRQAPGKGLEWIGVIGINGATYYASWAKGRFTISKTSSTTVDLKMTSLTTEDTATYFCARGDIWG PGTLVTVSS.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 34) QSLEESGGRLVTPGTPLTLTCSVSGIDLSGYYMNWVRQAPGKGLEWIGVIGINGATYYASWAKGRFTISKTSSTTVDLKMTSLTTEDTATYFCARGDIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 35; SEQ ID NO: 36; and SEQ IDNO: 37 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 31 or the light chain sequence of SEQ ID NO: 32, and/or oneor more of the polypeptide sequences of SEQ ID NO: 38; SEQ ID NO: 39;and SEQ ID NO: 40 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 33 or the heavy chain sequence of SEQ ID NO: 34,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 31 or SEQ ID NO: 32. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 33 or SEQ ID NO: 34.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 35; SEQ IDNO: 36; and SEQ ID NO: 37 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 31 or the light chainsequence of SEQ ID NO: 32.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 38; SEQ IDNO: 39; and SEQ ID NO: 40 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 33 or the heavy chainsequence of SEQ ID NO: 34.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 31; the variable heavy chain region of SEQ IDNO: 33; the complementarity-determining regions (SEQ ID NO: 35; SEQ IDNO: 36; and SEQ ID NO: 37) of the variable light chain region of SEQ IDNO: 31; and the complementarity-determining regions (SEQ ID NO: 38; SEQID NO: 39; and SEQ ID NO: 40) of the variable heavy chain region of SEQID NO: 33.

In a particularly preferred embodiment of the invention, the humanizedanti-CGRP antibody is Ab4, comprising, or alternatively consisting of,SEQ ID NO: 32 and SEQ ID NO: 34, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab4, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 31 and the variable heavy chain sequence ofSEQ ID NO: 33. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 31 and/or SEQ ID NO: 33in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab4.In another embodiment of the invention, anti-CGRP antibodies such as Ab4or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Ab5

In one embodiment, the invention includes humanized antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 41) QVLTQSPSSLSASVGDRVTINCQASQSVYHNTYLAWYQQKPGKVPKQLIYDASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCTNGDC FVFGGGTKVEIKR.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 42) QVLTQSPSSLSASVGDRVTINCQASQSVYHNTYLAWYQQKPGKVPKQLIYDASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCTNGDCFVFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC.

The invention further includes humanized antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 43) EVQLVESGGGLVQPGGSLRLSCAVSGIDLSGYYMNWVRQAPGKGLEWVGVIGINGATYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARGDI WGQGTLVTVSS.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 44) EVQLVESGGGLVQPGGSLRLSCAVSGIDLSGYYMNWVRQAPGKGLEWVGVIGINGATYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARGDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 45; SEQ ID NO: 46; and SEQ IDNO: 47 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 41 or the light chain sequence of SEQ ID NO: 42, and/or oneor more of the polypeptide sequences of SEQ ID NO: 48; SEQ ID NO: 49;and SEQ ID NO: 50 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 43 or the heavy chain sequence of SEQ ID NO: 44,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 41 or SEQ ID NO: 42. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 43 or SEQ ID NO: 44.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 45; SEQ IDNO: 46; and SEQ ID NO: 47 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 41 or the light chainsequence of SEQ ID NO: 42.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 48; SEQ IDNO: 49; and SEQ ID NO: 50 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 43 or the heavy chainsequence of SEQ ID NO: 44.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 41; the variable heavy chain region of SEQ IDNO: 43; the complementarity-determining regions (SEQ ID NO: 45; SEQ IDNO: 46; and SEQ ID NO: 47) of the variable light chain region of SEQ IDNO: 41; and the complementarity-determining regions (SEQ ID NO: 48; SEQID NO: 49; and SEQ ID NO: 50) of the variable heavy chain region of SEQID NO: 43.

In a particularly preferred embodiment of the invention, the chimericanti-CGRP antibody is Ab5, comprising, or alternatively consisting of,SEQ ID NO: 42 and SEQ ID NO: 44, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab5, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 41 and the variable heavy chain sequence ofSEQ ID NO: 43. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 41 and/or SEQ ID NO: 43in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab5.In another embodiment of the invention, anti-CGRP antibodies such asAb5or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Ab6

In one embodiment, the invention includes humanized antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 51) QVLTQSPSSLSASVGDRVTINCQASQSVYHNTYLAWYQQKPGKVPKQLIYDASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCTNGDC FVFGGGTKVEIKR.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 52) QVLTQSPSSLSASVGDRVTINCQASQSVYHNTYLAWYQQKPGKVPKQLIYDASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCTNGDCFVFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC.

The invention further includes humanized antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 53) EVQLVESGGGLVQPGGSLRLSCAVSGIDLSGYYMNWVRQAPGKGLEWVGVIGINGATYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARGDI WGQGTLVTVSS.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 54) EVQLVESGGGLVQPGGSLRLSCAVSGIDLSGYYMNWVRQAPGKGLEWVGVIGINGATYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARGDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56; and SEQ IDNO: 57 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 51 or the light chain sequence of SEQ ID NO: 52, and/or oneor more of the polypeptide sequences of SEQ ID NO: 58; SEQ ID NO: 59;and SEQ ID NO: 60 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 53 or the heavy chain sequence of SEQ ID NO: 54,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 51 or SEQ ID NO: 52. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 53 or SEQ ID NO: 54.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 55; SEQ IDNO: 56; and SEQ ID NO: 57 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 51 or the light chainsequence of SEQ ID NO: 52.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 58; SEQ IDNO: 59; and SEQ ID NO: 60 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 53 or the heavy chainsequence of SEQ ID NO: 54.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 51; the variable heavy chain region of SEQ IDNO: 53; the complementarity-determining regions (SEQ ID NO: 55; SEQ IDNO: 56; and SEQ ID NO: 57) of the variable light chain region of SEQ IDNO: 51; and the complementarity-determining regions (SEQ ID NO: 58; SEQID NO: 59; and SEQ ID NO: 60) of the variable heavy chain region of SEQID NO: 53.

In a particularly preferred embodiment of the invention, the humanizedanti-CGRP antibody is Ab6, comprising, or alternatively consisting of,SEQ ID NO: 52 and SEQ ID NO: 54, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab6, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 51 and the variable heavy chain sequence ofSEQ ID NO: 53. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 51 and/or SEQ ID NO: 53in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab6.In another embodiment of the invention, anti-CGRP antibodies such as Ab6or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Ab7

In one embodiment, the invention includes chimeric antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 61) QVLTQTASPVSAAVGSTVTINCQASQSVYNYNYLAWYQQKPGQPPKQLIYSTSTLASGVSSRFKGSGSGTQFTLTISDVQCDDAATYYCLGSYDCSTG DCFVFGGGTEVVVKR.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 62) QVLTQTASPVSAAVGSTVTINCQASQSVYNYNYLAWYQQKPGQPPKQLIYSTSTLASGVSSRFKGSGSGTQFTLTISDVQCDDAATYYCLGSYDCSTGDCFVFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 63) QEQLKESGGRLVTPGTSLTLTCTVSGIDLSNHYMQWVRQAPGKGLEWIGVVGINGRTYYASWAKGRFTISRTSSTTVDLKMTRLTTEDTATYFCARGD IWGPGTLVTVSS.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 64) QEQLKESGGRLVTPGTSLTLTCTVSGIDLSNHYMQWVRQAPGKGLEWIGVVGINGRTYYASWAKGRFTISRTSSTTVDLKMTRLTTEDTATYFCARGDIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 65; SEQ ID NO: 66; and SEQ IDNO: 67 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 61 or the light chain sequence of SEQ ID NO: 62, and/or oneor more of the polypeptide sequences of SEQ ID NO: 68; SEQ ID NO: 69;and SEQ ID NO: 70 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 63 or the heavy chain sequence of SEQ ID NO: 64,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 61 or SEQ ID NO: 62. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 63 or SEQ ID NO: 64.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 65; SEQ IDNO: 66; and SEQ ID NO: 67 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 61 or the light chainsequence of SEQ ID NO: 62.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 68; SEQ IDNO: 69; and SEQ ID NO: 70 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 63 or the heavy chainsequence of SEQ ID NO: 64.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 61; the variable heavy chain region of SEQ IDNO: 63; the complementarity-determining regions (SEQ ID NO: 65; SEQ IDNO: 66; and SEQ ID NO: 67) of the variable light chain region of SEQ IDNO: 61; and the complementarity-determining regions (SEQ ID NO: 68; SEQID NO: 69; and SEQ ID NO: 70) of the variable heavy chain region of SEQID NO: 63.

In a particularly preferred embodiment of the invention, the chimericanti-CGRP antibody is Ab7, comprising, or alternatively consisting of,SEQ ID NO: 62 and SEQ ID NO: 64, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab7, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 61 and the variable heavy chain sequence ofSEQ ID NO: 63. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 61 and/or SEQ ID NO: 63in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab7.In another embodiment of the invention, anti-CGRP antibodies such as Ab7or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Ab8

In one embodiment, the invention includes humanized antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 71) QVLTQSPSSLSASVGDRVTINCQASQSVYNYNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSTG DCFVFGGGTKVEIKR.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 72) QVLTQSPSSLSASVGDRVTINCQASQSVYNYNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSTGDCFVFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

The invention further includes humanized antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 73) EVQLVESGGGLVQPGGSLRLSCAVSGIDLSNHYMQWVRQAPGKGLEWVGVVGINGRTYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARG DIWGQGTLVTVSS.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 74) EVQLVESGGGLVQPGGSLRLSCAVSGIDLSNHYMQWVRQAPGKGLEWVGVVGINGRTYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARGDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 75; SEQ ID NO: 76; and SEQ IDNO: 77 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 71 or the light chain sequence of SEQ ID NO: 72, and/or oneor more of the polypeptide sequences of SEQ ID NO: 78; SEQ ID NO: 79;and SEQ ID NO: 80 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 73 or the heavy chain sequence of SEQ ID NO: 74,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 71 or SEQ ID NO: 72. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 73 or SEQ ID NO: 74.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 75; SEQ IDNO: 76; and SEQ ID NO: 77 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 71 or the light chainsequence of SEQ ID NO: 72.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 78; SEQ IDNO: 79; and SEQ ID NO: 80 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 73 or the heavy chainsequence of SEQ ID NO: 74.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 71; the variable heavy chain region of SEQ IDNO: 73; the complementarity-determining regions (SEQ ID NO: 75; SEQ IDNO: 76; and SEQ ID NO: 77) of the variable light chain region of SEQ IDNO: 71; and the complementarity-determining regions (SEQ ID NO: 78; SEQID NO: 79; and SEQ ID NO: 80) of the variable heavy chain region of SEQID NO: 73.

In a particularly preferred embodiment of the invention, the humanizedanti-CGRP antibody is Ab8, comprising, or alternatively consisting of,SEQ ID NO: 72 and SEQ ID NO: 74, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab8, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 71 and the variable heavy chain sequence ofSEQ ID NO: 73. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 71 and/or SEQ ID NO: 73in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab8.In another embodiment of the invention, anti-CGRP antibodies such as Ab8or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Ab9

In one embodiment, the invention includes chimeric antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 81) QVLTQTPSPVSAAVGSTVTINCQASQNVYNNNYLAWYQQKPGQPPKQLIYSTSTLASGVSSRFRGSGSGTQFTLTISDVQCDDAATYYCLGSYDCSRG DCFVFGGGTEVVVKR.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 82) QVLTQTPSPVSAAVGSTVTINCQASQNVYNNNYLAWYQQKPGQPPKQLIYSTSTLASGVSSRFRGSGSGTQFTLTISDVQCDDAATYYCLGSYDCSRGDCFVFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

The invention further includes chimeric antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 83) QSLEESGGRLVTPGTPLTLTCTVSGIGLSSYYMQWVRQSPGRGLEWIGVIGSDGKTYYATWAKGRFTISKTSSTTVDLRMASLTTEDTATYFCTRGDI WGPGTLVTVSS.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 84) QSLEESGGRLVTPGTPLTLTCTVSGIGLSSYYMQWVRQSPGRGLEWIGVIGSDGKTYYATWAKGRFTISKTSSTTVDLRMASLTTEDTATYFCTRGDIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 85; SEQ ID NO: 86; and SEQ IDNO: 87 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 81 or the light chain sequence of SEQ ID NO: 82, and/or oneor more of the polypeptide sequences of SEQ ID NO: 88; SEQ ID NO: 89;and SEQ ID NO: 90 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 83 or the heavy chain sequence of SEQ ID NO: 84,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 81 or SEQ ID NO: 82. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 83 or SEQ ID NO: 84.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 85; SEQ IDNO: 86; and SEQ ID NO: 87 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 81 or the light chainsequence of SEQ ID NO: 82.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 88; SEQ IDNO: 89; and SEQ ID NO: 90 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 83 or the heavy chainsequence of SEQ ID NO: 84.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 81; the variable heavy chain region of SEQ IDNO: 83; the complementarity-determining regions (SEQ ID NO: 85; SEQ IDNO: 86; and SEQ ID NO: 87) of the variable light chain region of SEQ IDNO: 81; and the complementarity-determining regions (SEQ ID NO: 88; SEQID NO: 89; and SEQ ID NO: 90) of the variable heavy chain region of SEQID NO: 83.

In a particularly preferred embodiment of the invention, the chimericanti-CGRP antibody is Ab9, comprising, or alternatively consisting of,SEQ ID NO: 82 and SEQ ID NO: 84, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab9, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 81 and the variable heavy chain sequence ofSEQ ID NO: 83. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 81 and/or SEQ ID NO: 83in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab9.In another embodiment of the invention, anti-CGRP antibodies such as Ab9or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Ab10

In one embodiment, the invention includes humanized antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 91) QVLTQSPSSLSASVGDRVTINCQASQNVYNNNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSRG DCFVFGGGTKVEIKR.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 92) QVLTQSPSSLSASVGDRVTINCQASQNVYNNNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSRGDCFVFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

The invention further includes humanized antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 93) EVQLVESGGGLVQPGGSLRLSCAVSGIGLSSYYMQWVRQAPGKGLEWVGVIGSDGKTYYATWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCTRG DIWGQGTLVTVSS.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 94) EVQLVESGGGLVQPGGSLRLSCAVSGIGLSSYYMQWVRQAPGKGLEWVGVIGSDGKTYYATWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCTRGDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 95; SEQ ID NO: 96; and SEQ IDNO: 97 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 91 or the light chain sequence of SEQ ID NO: 92, and/or oneor more of the polypeptide sequences of SEQ ID NO: 98; SEQ ID NO: 99;and SEQ ID NO: 100 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 93 or the heavy chain sequence of SEQ ID NO: 94,or combinations of these polypeptide sequences. In another embodiment ofthe invention, the antibodies of the invention or fragments thereofcomprise, or alternatively consist of, combinations of one or more ofthe CDRs, the variable heavy and variable light chain sequences, and theheavy and light chain sequences set forth above, including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 91 or SEQ ID NO: 92. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 93 or SEQ ID NO: 94.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 95; SEQ IDNO: 96; and SEQ ID NO: 97 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 91 or the light chainsequence of SEQ ID NO: 92.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 98; SEQ IDNO: 99; and SEQ ID NO: 100 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 93 or the heavy chainsequence of SEQ ID NO: 94.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 91; the variable heavy chain region of SEQ IDNO: 93; the complementarity-determining regions (SEQ ID NO: 95; SEQ IDNO: 96; and SEQ ID NO: 97) of the variable light chain region of SEQ IDNO: 91; and the complementarity-determining regions (SEQ ID NO: 98; SEQID NO: 99; and SEQ ID NO: 100) of the variable heavy chain region of SEQID NO: 93.

In a particularly preferred embodiment of the invention, the humanizedanti-CGRP antibody is Ab10, comprising, or alternatively consisting of,SEQ ID NO: 92 and SEQ ID NO: 94, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab10, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 91 and the variable heavy chain sequence ofSEQ ID NO: 93. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 91 and/or SEQ ID NO: 93in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab10.In another embodiment of the invention, anti-CGRP antibodies such asAb10or Fab fragments thereof may be produced via expression in mammaliancells such as CHO, NSO or HEK 293 cells, fungal, insect, or microbialsystems such as yeast cells (for example diploid yeast such as diploidPichia) and other yeast strains. Suitable Pichia species include, butare not limited to, Pichia pastoris.

Antibody Ab11

In one embodiment, the invention includes chimeric antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 101) QVLTQTASPVSPAVGSTVTINCRASQSVYYNNYLAWYQQKPGQPPKQLIYSTSTLASGVSSRFKGSGSGTQFTLTISDVQCDDAATYYCLGSYDCSNG DCFVFGGGTEVVVKR.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 102) QVLTQTASPVSPAVGSTVTINCRASQSVYYNNYLAWYQQKPGQPPKQLIYSTSTLASGVSSRFKGSGSGTQFTLTISDVQCDDAATYYCLGSYDCSNGDCFVFGGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 103) QSLEESGGRLVTPGGSLTLTCTVSGIDVTNYYMQWVRQAPGKGLEWIGVIGVNGKRYYASWAKGRFTISKTSSTTVDLKMTSLTTEDTATYFCARGDI WGPGTLVTVSS.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 104) QSLEESGGRLVTPGGSLTLTCTVSGIDVTNYYMQWVRQAPGKGLEWIGVIGVNGKRYYASWAKGRFTISKTSSTTVDLKMTSLTTEDTATYFCARGDIWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 105; SEQ ID NO: 106; and SEQ IDNO: 107 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 101 or the light chain sequence of SEQ ID NO: 102, and/or oneor more of the polypeptide sequences of SEQ ID NO: 108; SEQ ID NO: 109;and SEQ ID NO: 110 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 103 or the heavy chain sequence of SEQ ID NO:104, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the CDRs, the variable heavy and variable light chainsequences, and the heavy and light chain sequences set forth above,including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 101 or SEQ ID NO: 102. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 103 or SEQ ID NO: 104.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 105; SEQ IDNO: 106; and SEQ ID NO: 107 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 101 or the light chainsequence of SEQ ID NO: 102.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 108; SEQ IDNO: 109; and SEQ ID NO: 110 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 103 or the heavy chainsequence of SEQ ID NO: 104.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 101; the variable heavy chain region of SEQID NO: 103; the complementarity-determining regions (SEQ ID NO: 105; SEQID NO: 106; and SEQ ID NO: 107) of the variable light chain region ofSEQ ID NO: 101; and the complementarity-determining regions (SEQ ID NO:108; SEQ ID NO: 109; and SEQ ID NO: 110) of the variable heavy chainregion of SEQ ID NO: 103.

In a particularly preferred embodiment of the invention, the chimericanti-CGRP antibody is Ab11, comprising, or alternatively consisting of,SEQ ID NO: 102 and SEQ ID NO: 104, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab11, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 101 and the variable heavy chain sequenceof SEQ ID NO: 103. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 101 and/or SEQ ID NO:103 in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab11.In another embodiment of the invention, anti-CGRP antibodies such asAb11 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab12

In one embodiment, the invention includes humanized antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 111) QVLTQSPSSLSASVGDRVTINCRASQSVYYNNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSNG DCFVFGGGTKVEIKR.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 112) QVLTQSPSSLSASVGDRVTINCRASQSVYYNNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSNGDCFVFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

The invention further includes humanized antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 113) EVQLVESGGGLVQPGGSLRLSCAVSGIDVTNYYMQWVRQAPGKGLEWVGVIGVNGKRYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARG DIWGQGTLVTVSS.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 114) EVQLVESGGGLVQPGGSLRLSCAVSGIDVTNYYMQWVRQAPGKGLEWVGVIGVNGKRYYASWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCARGDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 115; SEQ ID NO: 116; and SEQ IDNO: 117 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 111 or the light chain sequence of SEQ ID NO: 112, and/or oneor more of the polypeptide sequences of SEQ ID NO: 118; SEQ ID NO: 119;and SEQ ID NO: 120 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 113 or the heavy chain sequence of SEQ ID NO:114, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the CDRs, the variable heavy and variable light chainsequences, and the heavy and light chain sequences set forth above,including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 111 or SEQ ID NO: 112. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 113 or SEQ ID NO: 114.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 115; SEQ IDNO: 116; and SEQ ID NO: 117 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 111 or the light chainsequence of SEQ ID NO: 112.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 118; SEQ IDNO: 119; and SEQ ID NO: 120 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 113 or the heavy chainsequence of SEQ ID NO: 114.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 111; the variable heavy chain region of SEQID NO: 113; the complementarity-determining regions (SEQ ID NO: 115; SEQID NO: 116; and SEQ ID NO: 117) of the variable light chain region ofSEQ ID NO: 111; and the complementarity-determining regions (SEQ ID NO:118; SEQ ID NO: 119; and SEQ ID NO: 120) of the variable heavy chainregion of SEQ ID NO: 113.

In a particularly preferred embodiment of the invention, the humanizedanti-CGRP antibody is Ab12, comprising, or alternatively consisting of,SEQ ID NO: 112 and SEQ ID NO: 114, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab12, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 111 and the variable heavy chain sequenceof SEQ ID NO: 113. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 111 and/or SEQ ID NO:113 in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab12.In another embodiment of the invention, anti-CGRP antibodies such asAb12 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab13

In one embodiment, the invention includes chimeric antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 121) AIVMTQTPSSKSVPVGDTVTINCQASESLYNNNALAWFQQKPGQPPKRLIYDASKLASGVPSRFSGGGSGTQFTLTISGVQCDDAATYYCGGYRSDSV DGVAFAGGTEVVVKR.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 122) AIVMTQTPSSKSVPVGDTVTINCQASESLYNNNALAWFQQKPGQPPKRLIYDASKLASGVPSRFSGGGSGTQFTLTISGVQCDDAATYYCGGYRSDSVDGVAFAGGTEVVVKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

The invention further includes chimeric antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 123) QSVEESGGGLVQPEGSLTLTCTASGFDFSSNAMWWVRQAPGKGLEWIGIIYNGDGSTYYASWVNGRFSISKTSSTTVTLQLNSLTVADTATYYCARDL DLWGPGTLVTVSS.

The invention also includes chimeric antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 124) QSVEESGGGLVQPEGSLTLTCTASGFDFSSNAMWWVRQAPGKGLEWIGCIYNGDGSTYYASWVNGRFSISKTSSTTVTLQLNSLTVADTATYYCARDLDLWGPGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 125; SEQ ID NO: 126; and SEQ IDNO: 127 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 121 or the light chain sequence of SEQ ID NO: 122, and/or oneor more of the polypeptide sequences of SEQ ID NO: 128; SEQ ID NO: 129;and SEQ ID NO: 130 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 123 or the heavy chain sequence of SEQ ID NO:124, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the CDRs, the variable heavy and variable light chainsequences, and the heavy and light chain sequences set forth above,including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 121 or SEQ ID NO: 122. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 123 or SEQ ID NO: 124.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 125; SEQ IDNO: 126; and SEQ ID NO: 127 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 121 or the light chainsequence of SEQ ID NO: 122.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 128; SEQ IDNO: 129; and SEQ ID NO: 130 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 123 or the heavy chainsequence of SEQ ID NO: 124.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 121; the variable heavy chain region of SEQID NO: 123; the complementarity-determining regions (SEQ ID NO: 125; SEQID NO: 126; and SEQ ID NO: 127) of the variable light chain region ofSEQ ID NO: 121; and the complementarity-determining regions (SEQ ID NO:128; SEQ ID NO: 129; and SEQ ID NO: 130) of the variable heavy chainregion of SEQ ID NO: 123.

In a particularly preferred embodiment of the invention, the chimericanti-CGRP antibody is Ab13, comprising, or alternatively consisting of,SEQ ID NO: 122 and SEQ ID NO: 124, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab13, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 121 and the variable heavy chain sequenceof SEQ ID NO: 123. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 121 and/or SEQ ID NO:123 in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab13.In another embodiment of the invention, anti-CGRP antibodies such asAb13 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab14

In one embodiment, the invention includes humanized antibodies havingbinding specificity to CGRP and possessing a variable light chainsequence comprising the sequence set forth below:

(SEQ ID NO: 131) QVLTQSPSSLSASVGDRVTINCQASQNVYNNNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSRG DCFVFGGGTKVEIKR.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a light chain sequence comprising thesequence set forth below:

(SEQ ID NO: 132) QVLTQSPSSLSASVGDRVTINCQASQNVYNNNYLAWYQQKPGKVPKQLIYSTSTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCLGSYDCSRGDCFVFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC.

The invention further includes humanized antibodies having bindingspecificity to CGRP and possessing a variable heavy chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 133) EVQLVESGGGLVQPGGSLRLSCAVSGIGLSSYYMQWVRQAPGKGLEWVGVIGSDGKTYYATWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCTRGDI WGQGTLVTVSS.

The invention also includes humanized antibodies having bindingspecificity to CGRP and possessing a heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 134) EVQLVESGGGLVQPGGSLRLSCAVSGIGLSSYYMQWVRQAPGKGLEWVGVIGSDGKTYYATWAKGRFTISRDNSKTTVYLQMNSLRAEDTAVYFCTRGDIWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDARVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 135; SEQ ID NO: 136; and SEQ IDNO: 137 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 131 or the light chain sequence of SEQ ID NO: 132, and/or oneor more of the polypeptide sequences of SEQ ID NO: 138; SEQ ID NO: 139;and SEQ ID NO: 140 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 133 or the heavy chain sequence of SEQ ID NO:134, or combinations of these polypeptide sequences. In anotherembodiment of the invention, the antibodies of the invention orfragments thereof comprise, or alternatively consist of, combinations ofone or more of the CDRs, the variable heavy and variable light chainsequences, and the heavy and light chain sequences set forth above,including all of them.

The invention also contemplates fragments of the antibody having bindingspecificity to CGRP. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 131 or SEQ ID NO: 132. In anotherembodiment of the invention, antibody fragments of the inventioncomprise, or alternatively consist of, the polypeptide sequence of SEQID NO: 133 or SEQ ID NO: 134.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 135; SEQ IDNO: 136; and SEQ ID NO: 137 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 131 or the light chainsequence of SEQ ID NO: 132.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to CGRP comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 138; SEQ IDNO: 139; and SEQ ID NO: 140 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 133 or the heavy chainsequence of SEQ ID NO: 134.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toCGRP comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 131; the variable heavy chain region of SEQID NO: 133; the complementarity-determining regions (SEQ ID NO: 135; SEQID NO: 136; and SEQ ID NO: 137) of the variable light chain region ofSEQ ID NO: 131; and the complementarity-determining regions (SEQ ID NO:138; SEQ ID NO: 139; and SEQ ID NO: 140) of the variable heavy chainregion of SEQ ID NO: 133.

In a particularly preferred embodiment of the invention, the humanizedanti-CGRP antibody is Ab14, comprising, or alternatively consisting of,SEQ ID NO: 132 and SEQ ID NO: 134, and having at least one of thebiological activities set forth herein.

In a further particularly preferred embodiment of the invention,antibody fragments comprise, or alternatively consist of, Fab (fragmentantigen binding) fragments having binding specificity for CGRP. Withrespect to antibody Ab14, the Fab fragment includes the variable lightchain sequence of SEQ ID NO: 131 and the variable heavy chain sequenceof SEQ ID NO: 133. This embodiment of the invention further contemplatesadditions, deletions, and variants of SEQ ID NO: 131 and/or SEQ ID NO:133 in said Fab while retaining binding specificity for CGRP.

In one embodiment of the invention described herein (infra), Fabfragments may be produced by enzymatic digestion (e.g., papain) of Ab14.In another embodiment of the invention, anti-CGRP antibodies such asAb14 or Fab fragments thereof may be produced via expression inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

In another embodiment, antibody fragments may be present in one or moreof the following non-limiting forms: Fab, Fab′, F(ab′)₂, Fv and singlechain Fv antibody forms. In a preferred embodiment, the anti-CGRPantibodies described herein further comprises the kappa constant lightchain sequence comprising the sequence set forth below:

(SEQ ID NO: 283) VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGEC.

In another preferred embodiment, the anti-CGRP antibodies describedherein further comprises the gamma-1 constant heavy chain polypeptidesequence comprising the sequence set forth below:

(SEQ ID NO: 284) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention contemplates an isolated anti-CGRPantibody comprising a V_(H) polypeptide sequence selected from: SEQ IDNO: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, or 133, or avariant thereof; and further comprising a V_(L) polypeptide sequenceselected from: SEQ ID NO: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101,111, 121, or 131, or a variant thereof, wherein one or more of theframework residues (FR residues) in said V_(H or) V_(L) polypeptide hasbeen substituted with another amino acid residue resulting in ananti-CGRP antibody that specifically binds CGRP. The inventioncontemplates humanized and chimeric forms of these antibodies. Thechimeric antibodies may include an Fc derived from IgG1, IgG2, IgG3,IgG4, IgG5, IgG6, IgG7, IgG8, IgG9, IgG10, IgG11, IgG12, IgG13, IgG14,IgG15, IgG16, IgG17, IgG18 or IgG19 constant regions.

In one embodiment of the invention, the antibodies or V_(H) or V_(L)polypeptides originate or are selected from one or more rabbit B cellpopulations prior to initiation of the humanization process referencedherein.

In another embodiment of the invention, the anti-CGRP antibodies andfragments thereof do not have binding specificity for CGRP-R. In afurther embodiment of the invention, the anti-CGRP antibodies andfragments thereof inhibit the association of CGRP with CGRP-R. Inanother embodiment of the invention, the anti-CGRP antibodies andfragments thereof inhibit the association of CGRP with CGRP-R and/oradditional proteins and/or multimers thereof, and/or antagonizes thebiological effects thereof.

As stated in paragraph [0127] herein, antibodies and fragments thereofmay be modified post-translationally to add effector moieties such aschemical linkers, detectable moieties such as for example fluorescentdyes, enzymes, substrates, bioluminescent materials, radioactivematerials, and chemiluminescent moieties, or functional moieties such asfor example streptavidin, avidin, biotin, a cytotoxin, a cytotoxicagent, and radioactive materials.

Antibodies or fragments thereof may also be chemically modified toprovide additional advantages such as increased solubility, stabilityand circulating time (in vivo half-life) of the polypeptide, ordecreased immunogenicity (See U.S. Pat. No. 4,179,337). The chemicalmoieties for derivatization may be selected from water soluble polymerssuch as polyethylene glycol, ethylene glycol/propylene glycolcopolymers, carboxymethylcellulose, dextran, polyvinyl alcohol and thelike. The antibodies and fragments thereof may be modified at randompositions within the molecule, or at predetermined positions within themolecule and may include one, two, three or more attached chemicalmoieties.

The polymer may be of any molecular weight, and may be branched orunbranched. For polyethylene glycol, the preferred molecular weight isbetween about 1 kDa and about 100 kDa (the term “about” indicating thatin preparations of polyethylene glycol, some molecules will weigh more,some less, than the stated molecular weight) for ease in handling andmanufacturing. Other sizes may be used, depending on the desiredtherapeutic profile (e.g., the duration of sustained release desired,the effects, if any on biological activity, the ease in handling, thedegree or lack of antigenicity and other known effects of thepolyethylene glycol to a therapeutic protein or analog). For example,the polyethylene glycol may have an average molecular weight of about200, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500,6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000,11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500,16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, 20,000,25,000, 30,000, 35,000, 40,000, 50,000, 55,000, 60,000, 65,000, 70,000,75,000, 80,000, 85,000, 90,000, 95,000, or 100,000 kDa. Branchedpolyethylene glycols are described, for example, in U.S. Pat. No.5,643,575; Morpurgo et al., Appl. Biochem. Biotechnol. 56:59-72 (1996);Vorobjev et al., Nucleosides Nucleotides 18:2745-2750 (1999); andCaliceti et al., Bioconjug. Chem. 10:638-646 (1999), the disclosures ofeach of which are incorporated herein by reference.

There are a number of attachment methods available to those skilled inthe art, See e.g., EP 0 401 384, herein incorporated by reference(coupling PEG to G-CSF), See also Malik et al., Exp. Hematol.20:1028-1035 (1992) (reporting pegylation of GM-CSF using tresylchloride). For example, polyethylene glycol may be covalently boundthrough amino acid residues via a reactive group, such as, a free aminoor carboxyl group. Reactive groups are those to which an activatedpolyethylene glycol molecule may be bound. The amino acid residueshaving a free amino group may include lysine residues and the N-terminalamino acid residues; those having a free carboxyl group may includeaspartic acid residues glutamic acid residues and the C-terminal aminoacid residue. Sulfhydryl groups may also be used as a reactive group forattaching the polyethylene glycol molecules. Preferred for therapeuticpurposes is attachment at an amino group, such as attachment at theN-terminus or lysine group.

As suggested above, polyethylene glycol may be attached to proteins vialinkage to any of a number of amino acid residues. For example,polyethylene glycol can be linked to polypeptides via covalent bonds tolysine, histidine, aspartic acid, glutamic acid, or cysteine residues.One or more reaction chemistries may be employed to attach polyethyleneglycol to specific amino acid residues (e.g., lysine, histidine,aspartic acid, glutamic acid, or cysteine) or to more than one type ofamino acid residue (e.g., lysine, histidine, aspartic acid, glutamicacid, cysteine and combinations thereof).

Alternatively, antibodies or fragments thereof may have increased invivo half lives via fusion with albumin (including but not limited torecombinant human serum albumin or fragments or variants thereof (See,e.g., U.S. Pat. No. 5,876,969, issued Mar. 2, 1999, EP Patent 0 413 622,and U.S. Pat. No. 5,766,883, issued Jun. 16, 1998, herein incorporatedby reference in their entirety)) or other circulating blood proteinssuch as transferrin or ferritin. In a preferred embodiment, polypeptidesand/or antibodies of the present invention (including fragments orvariants thereof) are fused with the mature form of human serum albumin(i.e., amino acids 1-585 of human serum albumin as shown in FIGS. 1 and2 of EP Patent 0 322 094) which is herein incorporated by reference inits entirety. Polynucleotides encoding fusion proteins of the inventionare also encompassed by the invention.

Regarding detectable moieties, further exemplary enzymes include, butare not limited to, horseradish peroxidase, acetylcholinesterase,alkaline phosphatase, beta-galactosidase and luciferase. Furtherexemplary fluorescent materials include, but are not limited to,rhodamine, fluorescein, fluorescein isothiocyanate, umbelliferone,dichlorotriazinylamine, phycoerythrin and dansyl chloride. Furtherexemplary chemiluminescent moieties include, but are not limited to,luminol. Further exemplary bioluminescent materials include, but are notlimited to, luciferin and aequorin. Further exemplary radioactivematerials include, but are not limited to, Iodine 125 (¹²⁵I Carbon 14(¹⁴C), Sulfur 35 (³⁵S), Tritium (³H) and Phosphorus 32 (³²P).

Regarding functional moieties, exemplary cytotoxic agents include, butare not limited to, methotrexate, aminopterin, 6-mercaptopurine,6-thioguanine, cytarabine, 5-fluorouracil decarbazine; alkylating agentssuch as mechlorethamine, thioepa chlorambucil, melphalan, carmustine(BSNU), mitomycin C, lomustine (CCNU), 1-methylnitrosourea,cyclothosphamide, mechlorethamine, busulfan, dibromomannitol,streptozotocin, mitomycin C, cis-dichlorodiamine platinum (II) (DDP)cisplatin and carboplatin (paraplatin); anthracyclines includedaunorubicin (formerly daunomycin), doxorubicin (adriamycin),detorubicin, caminomycin, idarubicin, epirubicin, mitoxantrone andbisantrene; antibiotics include dactinomycin (actinomycin D), bleomycin,calicheamicin, mithramycin, and anthramycin (AMC); and antimytoticagents such as the vinca alkaloids, vincristine and vinblastine. Othercytotoxic agents include paclitaxel (taxol), ricin, pseudomonasexotoxin, gemcitabine, cytochalasin B, gramicidin D, ethidium bromide,emetine, etoposide, tenoposide, colchicin, dihydroxy anthracin dione,1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine,propranolol, puromycin, procarbazine, hydroxyurea, asparaginase,corticosteroids, mytotane (O,P′-(DDD)), interferons, and mixtures ofthese cytotoxic agents.

Further cytotoxic agents include, but are not limited to,chemotherapeutic agents such as carboplatin, cisplatin, paclitaxel,gemcitabine, calicheamicin, doxorubicin, 5-fluorouracil, mitomycin C,actinomycin D, cyclophosphamide, vincristine and bleomycin. Toxicenzymes from plants and bacteria such as ricin, diphtheria toxin andPseudomonas toxin may be conjugated to the humanized or chimericantibodies, or binding fragments thereof, to generatecell-type-specific-killing reagents (Youle, et al., Proc. Nat'l Acad.Sci. USA 77:5483 (1980); Gilliland, et al., Proc. Nat'l Acad. Sci. USA77:4539 (1980); Krolick, et al., Proc. Nat'l Acad. Sci. USA 77:5419(1980)).

Other cytotoxic agents include cytotoxic ribonucleases as described byGoldenberg in U.S. Pat. No. 6,653,104. Embodiments of the invention alsorelate to radioimmunoconjugates where a radionuclide that emits alpha orbeta particles is stably coupled to the antibody, or binding fragmentsthereof, with or without the use of a complex-forming agent. Suchradionuclides include beta-emitters such as Phosphorus-32 (³²P),Scandium-47 (⁴⁷Sc), Copper-67 (⁶⁷Cu), Gallium-67 (⁶⁷Ga), Yttrium-88(⁸⁸Y), Yttrium-90 (⁹⁰Y), Iodine-125 (¹²⁵I) Iodine-131 (¹³¹I)Samarium-153 (¹⁵³Sm), Lutetium-177 (¹⁷⁷Lu), Rhenium-186 (¹⁸⁶Re) orRhenium-188 (¹⁸⁸Re), and alpha-emitters such as Astatine-211 (²¹¹At),Lead-212 (²¹²Pb), Bismuth-212 (²¹²Bi) or -213 (²¹³Bi) or Actinium-225(²²⁵Ac).

Methods are known in the art for conjugating an antibody or bindingfragment thereof to a detectable moiety and the like, such as forexample those methods described by Hunter et al, Nature 144:945 (1962);David et al, Biochemistry 13:1014 (1974); Pain et al, J. Immunol. Meth.40:219 (1981); and Nygren, J., Histochem. and Cytochem. 30:407 (1982).

Embodiments described herein further include variants and equivalentsthat are substantially homologous to the antibodies, antibody fragments,diabodies, SMIPs, camelbodies, nanobodies, IgNAR, polypeptides, variableregions and CDRs set forth herein. These may contain, e.g., conservativesubstitution mutations, (i.e., the substitution of one or more aminoacids by similar amino acids). For example, conservative substitutionrefers to the substitution of an amino acid with another within the samegeneral class, e.g., one acidic amino acid with another acidic aminoacid, one basic amino acid with another basic amino acid, or one neutralamino acid by another neutral amino acid. What is intended by aconservative amino acid substitution is well known in the art.

In another embodiment, the invention contemplates polypeptide sequenceshaving at least 90% or greater sequence homology to any one or more ofthe polypeptide sequences of antibody fragments, variable regions andCDRs set forth herein. More preferably, the invention contemplatespolypeptide sequences having at least 95% or greater sequence homology,even more preferably at least 98% or greater sequence homology, andstill more preferably at least 99% or greater sequence homology to anyone or more of the polypeptide sequences of antibody fragments, variableregions and CDRs set forth herein. Methods for determining homologybetween nucleic acid and amino acid sequences are well known to those ofordinary skill in the art.

In another embodiment, the invention further contemplates theabove-recited polypeptide homologs of the antibody fragments, variableregions and CDRs set forth herein further having anti-CGRP activity.Non-limiting examples of anti-CGRP activity are set forth herein, forexample, in paragraphs [0329]-[0350] infra.

In another embodiment, the invention further contemplates the generationand use of anti-idiotypic antibodies that bind any of the foregoingsequences. In an exemplary embodiment, such an anti-idiotypic antibodycould be administered to a subject who has received an anti-CGRPantibody to modulate, reduce, or neutralize, the effect of the anti-CGRPantibody. Such anti-idiotypic antibodies could also be useful fortreatment of an autoimmune disease characterized by the presence ofanti-CGRP antibodies. A further exemplary use of such anti-idiotypicantibodies is for detection of the anti-CGRP antibodies of the presentinvention, for example to monitor the levels of the anti-CGRP antibodiespresent in a subject's blood or other bodily fluids.

The present invention also contemplates anti-CGRP antibodies comprisingany of the polypeptide or polynucleotide sequences described hereinsubstituted for any of the other polynucleotide sequences describedherein. For example, without limitation thereto, the present inventioncontemplates antibodies comprising the combination of any of thevariable light chain and variable heavy chain sequences describedherein, and further contemplates antibodies resulting from substitutionof any of the CDR sequences described herein for any of the other CDRsequences described herein.

Additional Exemplary Embodiments of the Invention

In another embodiment, the invention contemplates one or more anti-humanCGRP antibodies or antibody fragments thereof which specifically bind tothe same ovelappping linear or conformational epitope(s) and/or competesfor binding to the same ovelappping linear or conformational epitope(s)on an intact human CGRP polypeptide or fragment thereof as an anti-humanCGRP antibody selected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9,Ab10, Ab11, Ab12, Ab13, or Ab14. In a preferred embodiment, theanti-human CGRP antibody or fragment thereof specifically binds to thesame ovelappping linear or conformational epitope(s) and/or competes forbinding to the same ovelappping linear or conformational epitope(s) onan intact human CGRP polypeptide or a fragment thereof as Ab3, Ab6,Ab13, or Ab14.

A preferred embodiment of the invention is directed to chimeric orhumanized antibodies and fragments thereof (including Fab fragments)having binding specificity for CGRP and inhibiting biological activitiesmediated by the binding of CGRP to the CGRP receptor. In a particularlypreferred embodiment of the invention, the chimeric or humanizedanti-CGRP antibodies are selected from Ab3, Ab6, Ab13, or Ab14.

In a further embodiment of the invention is contemplated a method ofreducing, treating or preventing diseases or disorders associated withCGRP by affecting those biological activities mediated via CGRP, therebyavoiding the biological activities mediated via binding of CGRP toCGRP-R. In one embodiment, the disease or disorder associated with CGRPis migraine or another disorder wherein CGRP elicits pain, headache,pain, cancer, overactive bladder, or weightloss. A further non-limitinglisting of diseases and disorders associated with CGRP is providedherein.

Another preferred embodiment of the invention contemplates the use ofFab polypeptide sequences for the treatment of migraines and headachesin a patient. Non-limiting types of migraines and headaches that may betreated using Fab polypeptide sequences are provided elsewhere in thisdisclosure.

In another embodiment of the invention, the anti-human CGRP antibody isan antibody which specifically binds to the same ovelappping linear orconformational epitopes on an intact CGRP polypeptide or fragmentthereof that is (are) specifically bound by Ab3, Ab6, Ab13, or Ab14 asascertained by epitopic mapping using overlapping linear peptidefragments which span the full length of the native human CGRPpolypeptide.

The invention is also directed to an anti-CGRP antibody that binds withthe same CGRP epitope and/or competes with an anti-CGRP antibody forbinding to CGRP as an antibody or antibody fragment disclosed herein,including but not limited to an anti-CGRP antibody selected from Ab1,Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, or Ab14.

In another embodiment, the invention is also directed to an isolatedanti-CGRP antibody or antibody fragment comprising one or more of theCDRs contained in the V_(H) polypeptide sequences selected from: 3, 13,23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, or 133, or a variantthereof, and/or one or more of the CDRs contained in the V_(L)polypeptide sequences selected from: 1, 11, 21, 31, 41, 51, 61, 71, 81,91, 101, 111, 121, or 131, or a variant thereof.

In one embodiment of the invention, the anti-human CGRP antibodydiscussed in the two prior paragraphs comprises at least 2complementarity determining regions (CDRs) in each the variable lightand the variable heavy regions which are identical to those contained inan anti-human CGRP antibody selected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, or Ab14.

In a preferred embodiment, the anti-human CGRP antibody discussed abovecomprises at least 2 complementarity determining regions (CDRs) in eachthe variable light and the variable heavy regions which are identical tothose contained in Ab3 or Ab6. In another embodiment, all of the CDRs ofthe anti-human CGRP antibody discussed above are identical to the CDRscontained in an anti-human CGRP antibody selected from Ab1, Ab2, Ab3,Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, or Ab14. In apreferred embodiment of the invention, all of the CDRs of the anti-humanCGRP antibody discussed above are identical to the CDRs contained in ananti-human CGRP antibody selected from Ab3 or Ab6.

The invention further contemplates that the one or more anti-human CGRPantibodies discussed above are aglycosylated; that contain an Fc regionthat has been modified to alter effector function, half-life,proteolysis, and/or glycosylation; are human, humanized, single chain orchimeric; and are a humanized antibody derived from a rabbit (parent)anti-human CGRP antibody.

The invention further contemplates one or more anti-human CGRPantibodies wherein the framework regions (FRs) in the variable lightregion and the variable heavy regions of said antibody respectively arehuman FRs which are unmodified or which have been modified by thesubstitution of one or more human FR residues in the variable light orheavy chain region with the corresponding FR residues of the parentrabbit antibody, and wherein said human FRs have been derived from humanvariable heavy and light chain antibody sequences which have beenselected from a library of human germline antibody sequences based ontheir high level of homology to the corresponding rabbit variable heavyor light chain regions relative to other human germline antibodysequences contained in the library.

In one embodiment of the invention, the anti-human CGRP antibody orfragment specifically binds to CGRP expressing human cells and/or tocirculating soluble CGRP molecules in vivo, including CGRP expressed onor by human cells in a patient with a disease associated with cells thatexpress CGRP.

In another embodiment, the disease is selected from migraines (with orwithout aura), weight loss, cancer or tumors, angiogenesis associatedwith cancer or tumor growth, angiogenesis associated with cancer ortumor survival, hemiplagic migraines, cluster headaches, migrainousneuralgia, chronic headaches, tension headaches, general headaches, hotflushes, chronic paroxysomal hemicrania, secondary headaches due to anunderlying structural problem in the head or neck, cranial neuralgia,sinus headaches (such as for example associated with sinusitis),allergy-induced headaches or migraines, pain, inflammatory pain,post-operative incision pain, complex regional pain syndrome, cancerpain, primary or metastatic bone cancer pain, fracture pain, chronicpain, osteoporotic fracture pain, pain resulting from burn,osteoporosis, gout joint pain, abdominal pain, pain associated withsickle cell crises, and other nociceptic pain, as well as hepatocellularcarcinoma, breast cancer, liver cirrhosis, neurogenic pain, neuropathicpain, nociceptic pain, trigeminal neuralgia, post-herpetic neuralgia,phantom limb pain, fibromyalgia, menstrual pain, ovarialgia, reflexsympathetic dystrophy, neurogenic pain, osteoarthritis or rheumatoidarthritis pain, lower back pain, diabetic neuropathy, sciatica, or painor visceral pain associated with: gastro-esophageal reflux, dyspepsia,irritable bowel syndrome, irritable colon, spastic colon, mucouscolitis, inflammatory bowel disease, Crohn's disease, ileitis,ulcerative colitis, renal colic, dysmenorrhea, cystitis, menstrualperiod, labor, menopause, prostatitis, pancreatitis, renal colic,dysmenorrhea, cystitis, including interstitial cystitis (IC), surgeryassociated with the ileus, diverticulitis, peritonitis, pericarditis,hepatitis, appendicitis, colitis, cholecystitis, endometriosis, chronicand/or acute pancreatitis, myocardial infarction, kidney pain, pleuralpain, prostatitis, pelvic pain, trauma to an organ, chronic nociceptivepain, chronic neuropathic pain, chronic inflammatory pain, fibromyalgia,breakthrough pain and persistent pain.

In another embodiment of the invention, the disease is cancer painarising from malignancy or from cancer preferably selected from one ormore of: adenocarcinoma in glandular tissue, blastoma in embryonictissue of organs, carcinoma in epithelial tissue, leukemia in tissuesthat form blood cells, lymphoma in lymphatic tissue, myeloma in bonemarrow, sarcoma in connective or supportive tissue, adrenal cancer,AIDS-related lymphoma, anemia, bladder cancer, bone cancer, braincancer, breast cancer, carcinoid tumours, cervical cancer, chemotherapy,colon cancer, cytopenia, endometrial cancer, esophageal cancer, gastriccancer, head cancer, neck cancer, hepatobiliary cancer, kidney cancer,leukemia, liver cancer, lung cancer, lymphoma, Hodgkin's disease,lymphoma, non-Hodgkin's, nervous system tumours, oral cancer, ovariancancer, pancreatic cancer, prostate cancer, rectal cancer, skin cancer,stomach cancer, testicular cancer, thyroid cancer, urethral cancer, bonecancer, sarcomas cancer of the connective tissue, cancer of bone tissue,cancer of blood-forming cells, cancer of bone marrow, multiple myeloma,leukaemia, primary or secondary bone cancer, tumours that metastasize tothe bone, tumours infiltrating the nerve and hollow viscus, tumours nearneural structures. Further preferably the cancer pain comprises visceralpain, preferably visceral pain which arises from pancreatic cancerand/or metastases in the abdomen. Further preferably the cancer paincomprises somatic pain, preferably somatic pain due to one or more ofbone cancer, metastasis in the bone, postsurgical pain, sarcomas cancerof the connective tissue, cancer of bone tissue, cancer of blood-formingcells of the bone marrow, multiple myeloma, leukaemia, primary orsecondary bone cancer.

The invention further contemplates anti-human CGRP antibodies orfragments directly or indirectly attached to a detectable label ortherapeutic agent.

The invention also contemplates one or more nucleic acid sequences whichresult in the expression of an anti-human CGRP antibody or antibodyfragment as set forth above, including those comprising, oralternatively consisting of, yeast or human preferred codons. Theinvention also contemplates vectors (including plasmids or recombinantviral vectors) comprising said nucleic acid sequence(s). The inventionalso contemplates host cells or recombinant host cells expressing atleast one of the antibodies set forth above, including a mammalian,yeast, bacterial, and insect cells. In a preferred embodiment, the hostcell is a yeast cell. In a further preferred embodiment, the yeast cellis a diploidal yeast cell. In a more preferred embodiment, the yeastcell is a Pichia yeast.

The invention also contemplates a method of treatment comprisingadministering to a patient with a disease or condition associated withCGRP expressing cells a therapeutically effective amount of at least oneanti-human CGRP antibody or fragment described herein. The inventionalso contemplates that the treatment method may involve theadministration of two or more anti-CGRP antibodies or fragments thereofand disclosed herein. If more than one antibody is administered to thepatient, the multiple antibodies may be administered simultaneously orconcurrently, or may be staggered in their administration. The diseasesthat may be treated are presented in the non-limiting list set forthabove and elsewhere herein. In a preferred embodiment, the disease isselected from migraine, headache, weight loss, pain, cancer pain orneuropathic pain. In another embodiment the treatment further includesthe administration of another therapeutic agent or regimen selected fromchemotherapy, radiotherapy, cytokine administration or gene therapy.

In a non-limiting embodiment of the invention, another therapeutic agentor regimen includes Taxol (paclitaxel) or its derivatives, platinumcompounds such as carboplatin or cisplatin, anthrocyclines such asdoxorubicin, alkylating agents such as cyclophosphamide,anti-metabolites such as 5-fluorouracil, or etoposide.

The invention further contemplates a method of in vivo imaging whichdetects the presence of cells which express CGRP comprisingadministering a diagnostically effective amount of at least oneanti-human CGRP antibody. In one embodiment, said administration furtherincludes the administration of a radionuclide or fluorophore thatfacilitates detection of the antibody at CGRP expressing disease sites.In a further embodiment, the results of said in vivo imaging method areused to facilitate the design of an appropriate therapeutic regimen,including therapeutic regimens including radiotherapy, chemotherapy or acombination thereof.

The anti-CGRP activity of the anti-CGRP antibodies of the presentinvention, and fragments thereof having binding specificity to CGRP, mayalso be described by their strength of binding or their affinity forCGRP. In one embodiment of the invention, the anti-CGRP antibodies ofthe present invention, and fragments thereof having binding specificityto CGRP, bind to CGRP with a dissociation constant (K_(D)) of less thanor equal to 5×10⁻⁷ M, 10⁻⁷ M, 5×10⁻⁸ M, 10⁻⁸ M, 5×10⁻⁹ M, 10⁻⁹ M,5×10⁻¹° M, 10⁻¹⁰ M, 5×10⁻¹¹ M, 10⁻¹¹ M, 5×10⁻¹² M, 10⁻¹² M, 5×10⁻¹³ M,or 10⁻¹³ M. Preferably, the anti-CGRP antibodies and fragments thereofbind CGRP with a dissociation constant of less than or equal to 10⁻¹¹ M,5×10⁻¹² M, or 10⁻¹² M. In another embodiment of the invention, theanti-CGRP antibodies of the present invention, and fragments thereofhaving binding specificity to CGRP, bind to a linear or conformationalCGRP epitope.

In another embodiment of the invention, the anti-CGRP activity of theanti-CGRP antibodies of the present invention, and fragments thereofhaving binding specificity to CGRP, bind to CGRP with an off-rate ofless than or equal to 10⁻⁴ S⁻¹, 5×10⁻⁵ S⁻¹, 10⁻⁵ S⁻¹, 5×10⁻⁶ S⁻¹, 10⁻⁶S⁻¹, 5×10⁻⁷ S⁻¹, or 10⁻⁷ S⁻¹.

In a further embodiment of the invention, the anti-CGRP activity of theanti-CGRP antibodies of the present invention, and fragments thereofhaving binding specificity to CGRP, exhibit anti-CGRP activity bypreventing, ameliorating or reducing the symptoms of, or alternativelytreating, diseases and disorders associated with CGRP. Non-limitingexamples of diseases and disorders associated with CGRP are set forthherein.

Polynucleotides Encoding Anti-CGRP Antibody Polypeptides Antibody Ab1

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 1:

(SEQ ID NO: 141) CAAGTGCTGACCCAGACTGCATCCCCCGTGTCTGCAGCTGTGGGAAGCACAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATGATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTACTACTGTCTAGGCAGTTATGATTGTAGTAGTGGTGATTGTTTTGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 2:

(SEQ ID NO: 142) CAAGTGCTGACCCAGACTGCATCCCCCGTGTCTGCAGCTGTGGGAAGCACAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATGATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTACTACTGTCTAGGCAGTTATGATTGTAGTAGTGGTGATTGTTTTGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGG AGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 3:

(SEQ ID NO: 143) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGACTCGACCTCAGTAGCTACTACATGCAATGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGTCATTGGTATTAATGATAACACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGCCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGGGACATCTGGGGCCCAGGCACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 4:

(SEQ ID NO: 144) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGACTCGACCTCAGTAGCTACTACATGCAATGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGTCATTGGTATTAATGATAACACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGCCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGGGACATCTGGGGCCCAGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCT CCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 145; SEQ ID NO: 146; and SEQ ID NO: 147 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 1 or the light chain sequence of SEQ ID NO: 2.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 148; SEQ ID NO: 149; and SEQ ID NO: 150 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 3 or the heavy chain sequence of SEQ ID NO: 4.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 141 encoding the light chain variable sequenceof SEQ ID NO: 1; the polynucleotide SEQ ID NO: 142 encoding the lightchain sequence of SEQ ID NO: 2; the polynucleotide SEQ ID NO: 143encoding the heavy chain variable sequence of SEQ ID NO: 3; thepolynucleotide SEQ ID NO: 144 encoding the heavy chain sequence of SEQID NO: 4; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 145; SEQ ID NO: 146; and SEQ ID NO: 147) of thelight chain variable sequence of SEQ ID NO: 1 or the light chainsequence of SEQ ID NO: 2; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 148; SEQ ID NO: 149; andSEQ ID NO: 150) of the heavy chain variable sequence of SEQ ID NO: 3 orthe heavy chain sequence of SEQ ID NO: 4.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab1, the polynucleotidesencoding the full length Ab1 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 142 encoding the light chain sequenceof SEQ ID NO: 2 and the polynucleotide SEQ ID NO: 144 encoding the heavychain sequence of SEQ ID NO: 4.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab1 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab1 or Fab fragments thereofmay be produced via expression of Ab1 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab2

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 11:

(SEQ ID NO: 151) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATGATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTAGGCAGTTATGATTGTAGTAGTGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 12:

(SEQ ID NO: 152) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATGATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTAGGCAGTTATGATTGTAGTAGTGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGG AGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 13:

(SEQ ID NO: 153) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGACTCGACCTCAGTAGCTACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTATCAATGATAACACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCTAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 14:

(SEQ ID NO: 154) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGACTCGACCTCAGTAGCTACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTATCAATGATAACACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCTAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 155; SEQ ID NO: 156; and SEQ ID NO: 157 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 11 or the light chain sequence of SEQ ID NO: 12.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 158; SEQ ID NO: 159; and SEQ ID NO: 160 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 13 or the heavy chain sequence of SEQ ID NO: 14.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 151 encoding the light chain variable sequenceof SEQ ID NO: 11; the polynucleotide SEQ ID NO: 152 encoding the lightchain sequence of SEQ ID NO: 12; the polynucleotide SEQ ID NO: 153encoding the heavy chain variable sequence of SEQ ID NO: 13; thepolynucleotide SEQ ID NO: 154 encoding the heavy chain sequence of SEQID NO: 14; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 155; SEQ ID NO: 156; and SEQ ID NO: 157) of thelight chain variable sequence of SEQ ID NO: 11 or the light chainsequence of SEQ ID NO: 12; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 158; SEQ ID NO: 159; andSEQ ID NO: 160) of the heavy chain variable sequence of SEQ ID NO: 13 orthe heavy chain sequence of SEQ ID NO: 14.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab2, the polynucleotidesencoding the full length Ab2 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 152 encoding the light chain sequenceof SEQ ID NO: 12 and the polynucleotide SEQ ID NO: 154 encoding theheavy chain sequence of SEQ ID NO: 14.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab2 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab2 or Fab fragments thereofmay be produced via expression of Ab2 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab3

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 21:

(SEQ ID NO: 161) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATGATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTAGGCAGTTATGATTGTAGTAGTGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 22:

(SEQ ID NO: 162) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATGATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTAGGCAGTTATGATTGTAGTAGTGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGG AGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 23:

(SEQ ID NO: 163) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGACTCGACCTCAGTAGCTACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTATCAATGATAACACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCTAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 24:

(SEQ ID NO: 164) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGACTCGACCTCAGTAGCTACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTATCAATGATAACACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCTAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACGCGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA TGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 165; SEQ ID NO: 166; and SEQ ID NO: 167 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 21 or the light chain sequence of SEQ ID NO: 22.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 168; SEQ ID NO: 169; and SEQ ID NO: 170 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 23 or the heavy chain sequence of SEQ ID NO: 24.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 161 encoding the light chain variable sequenceof SEQ ID NO: 21; the polynucleotide SEQ ID NO: 162 encoding the lightchain sequence of SEQ ID NO: 22; the polynucleotide SEQ ID NO: 163encoding the heavy chain variable sequence of SEQ ID NO: 23; thepolynucleotide SEQ ID NO: 164 encoding the heavy chain sequence of SEQID NO: 24; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 165; SEQ ID NO: 166; and SEQ ID NO: 167) of thelight chain variable sequence of SEQ ID NO: 21 or the light chainsequence of SEQ ID NO: 22; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 168; SEQ ID NO: 169; andSEQ ID NO: 170) of the heavy chain variable sequence of SEQ ID NO: 23 orthe heavy chain sequence of SEQ ID NO: 24.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab3, the polynucleotidesencoding the full length Ab3 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 162 encoding the light chain sequenceof SEQ ID NO: 22 and the polynucleotide SEQ ID NO: 164 encoding theheavy chain sequence of SEQ ID NO: 24.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab3 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab3 or Fab fragments thereofmay be produced via expression of Ab3 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab4

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 31:

(SEQ ID NO: 171) CAAGTGCTGACCCAGACTCCATCCCCCGTGTCTGCAGCTGTGGGAAGCACAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATCATAACACCTACCTGGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAACAACTGATCTATGATGCATCCACTCTGGCGTCTGGGGTCCCATCGCGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGTAACGATGCTGCCGCTTACTACTGTCTGGGCAGTTATGATTGTACTAATGGTGATTGTTTTGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 32:

(SEQ ID NO: 172) CAAGTGCTGACCCAGACTCCATCCCCCGTGTCTGCAGCTGTGGGAAGCACAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATCATAACACCTACCTGGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAACAACTGATCTATGATGCATCCACTCTGGCGTCTGGGGTCCCATCGCGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGTAACGATGCTGCCGCTTACTACTGTCTGGGCAGTTATGATTGTACTAATGGTGATTGTTTTGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 33:

(SEQ ID NO: 173) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGTTCCGTCTCTGGCATCGACCTCAGTGGCTACTACATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGTCATTGGTATTAATGGTGCCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGGGACATCTGGGGCCCGGGCACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 34:

(SEQ ID NO: 174) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGTTCCGTCTCTGGCATCGACCTCAGTGGCTACTACATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGTCATTGGTATTAATGGTGCCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGGGACATCTGGGGCCCGGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 175; SEQ ID NO: 176; and SEQ ID NO: 177 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 31 or the light chain sequence of SEQ ID NO: 32.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 178; SEQ ID NO: 179; and SEQ ID NO: 180 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 33 or the heavy chain sequence of SEQ ID NO: 34.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 171 encoding the light chain variable sequenceof SEQ ID NO: 31; the polynucleotide SEQ ID NO: 172 encoding the lightchain sequence of SEQ ID NO: 32; the polynucleotide SEQ ID NO: 173encoding the heavy chain variable sequence of SEQ ID NO: 33; thepolynucleotide SEQ ID NO: 174 encoding the heavy chain sequence of SEQID NO: 34; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 175; SEQ ID NO: 176; and SEQ ID NO: 177) of thelight chain variable sequence of SEQ ID NO: 31 or the light chainsequence of SEQ ID NO: 32; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 178; SEQ ID NO: 179; andSEQ ID NO: 180) of the heavy chain variable sequence of SEQ ID NO: 33 orthe heavy chain sequence of SEQ ID NO: 34.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab4, the polynucleotidesencoding the full length Ab4 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 172 encoding the light chain sequenceof SEQ ID NO: 32 and the polynucleotide SEQ ID NO: 174 encoding theheavy chain sequence of SEQ ID NO: 34.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab4 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab4 or Fab fragments thereofmay be produced via expression of Ab4 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab5

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 41:

(SEQ ID NO: 181) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATCATAACACCTACCTGGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATGATGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTACTAATGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 42:

(SEQ ID NO: 182) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATCATAACACCTACCTGGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATGATGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTACTAATGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 43:

(SEQ ID NO: 183) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGACCTCAGTGGCTACTACATGAACTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTATTAATGGTGCCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCTAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 44:

(SEQ ID NO: 184) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGACCTCAGTGGCTACTACATGAACTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTATTAATGGTGCCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCTAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA TGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 185; SEQ ID NO: 186; and SEQ ID NO: 187 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 41 or the light chain sequence of SEQ ID NO: 42.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 188; SEQ ID NO: 189; and SEQ ID NO: 190 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 43 or the heavy chain sequence of SEQ ID NO: 44.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 181 encoding the light chain variable sequenceof SEQ ID NO: 41; the polynucleotide SEQ ID NO: 182 encoding the lightchain sequence of SEQ ID NO: 42; the polynucleotide SEQ ID NO: 183encoding the heavy chain variable sequence of SEQ ID NO: 43; thepolynucleotide SEQ ID NO: 184 encoding the heavy chain sequence of SEQID NO: 44; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 185; SEQ ID NO: 186; and SEQ ID NO: 187) of thelight chain variable sequence of SEQ ID NO: 41 or the light chainsequence of SEQ ID NO: 42; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 188; SEQ ID NO: 189; andSEQ ID NO: 190) of the heavy chain variable sequence of SEQ ID NO: 43 orthe heavy chain sequence of SEQ ID NO: 44.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab5, the polynucleotidesencoding the full length Ab5 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 182 encoding the light chain sequenceof SEQ ID NO: 42 and the polynucleotide SEQ ID NO: 184 encoding theheavy chain sequence of SEQ ID NO: 44.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab5 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab5 or Fab fragments thereofmay be produced via expression of Ab5 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab6

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 51:

(SEQ ID NO: 191) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATCATAACACCTACCTGGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATGATGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTACTAATGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 52:

(SEQ ID NO: 192) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATCATAACACCTACCTGGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATGATGCATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTACTAATGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 53:

(SEQ ID NO: 193) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGACCTCAGTGGCTACTACATGAACTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTATTAATGGTGCCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCTAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 54:

(SEQ ID NO: 194) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGACCTCAGTGGCTACTACATGAACTGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTATTAATGGTGCCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCTAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACGCGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA TGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 195; SEQ ID NO: 196; and SEQ ID NO: 197 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 51 or the light chain sequence of SEQ ID NO: 52.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 198; SEQ ID NO: 199; and SEQ ID NO: 200 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 53 or the heavy chain sequence of SEQ ID NO: 54.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 191 encoding the light chain variable sequenceof SEQ ID NO: 51; the polynucleotide SEQ ID NO: 192 encoding the lightchain sequence of SEQ ID NO: 52; the polynucleotide SEQ ID NO: 193encoding the heavy chain variable sequence of SEQ ID NO: 53; thepolynucleotide SEQ ID NO: 194 encoding the heavy chain sequence of SEQID NO: 54; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 195; SEQ ID NO: 196; and SEQ ID NO: 197) of thelight chain variable sequence of SEQ ID NO: 51 or the light chainsequence of SEQ ID NO: 52; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 198; SEQ ID NO: 199; andSEQ ID NO: 200) of the heavy chain variable sequence of SEQ ID NO: 53 orthe heavy chain sequence of SEQ ID NO: 54.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab6, the polynucleotidesencoding the full length Ab6 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 192 encoding the light chain sequenceof SEQ ID NO: 52 and the polynucleotide SEQ ID NO: 194 encoding theheavy chain sequence of SEQ ID NO: 54.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab6 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab6 or Fab fragments thereofmay be produced via expression of Ab6 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab7

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 61:

(SEQ ID NO: 201) CAAGTGCTGACCCAGACTGCATCCCCCGTGTCTGCAGCTGTGGGAAGCACAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATAATTACAACTACCTTGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCTCATCGCGATTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCAGTTATGACTGTAGTACTGGTGATTGTTTTGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 62:

(SEQ ID NO: 202) CAAGTGCTGACCCAGACTGCATCCCCCGTGTCTGCAGCTGTGGGAAGCACAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATAATTACAACTACCTTGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCTCATCGCGATTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCAGTTATGACTGTAGTACTGGTGATTGTTTTGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 63:

(SEQ ID NO: 203) CAGGAGCAGCTGAAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACATCCCTGACACTCACCTGCACCGTCTCTGGAATCGACCTCAGTAACCACTACATGCAATGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCGTTGGTATTAATGGTCGCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAACCTCGTCGACCACGGTGGATCTGAAAATGACCAGGCTGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGGGACATCTGGGGCCCAGGCACCCTGGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 64:

(SEQ ID NO: 204) CAGGAGCAGCTGAAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACATCCCTGACACTCACCTGCACCGTCTCTGGAATCGACCTCAGTAACCACTACATGCAATGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCGTTGGTATTAATGGTCGCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAACCTCGTCGACCACGGTGGATCTGAAAATGACCAGGCTGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGGGACATCTGGGGCCCAGGCACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 205; SEQ ID NO: 206; and SEQ ID NO: 207 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 61 or the light chain sequence of SEQ ID NO: 62.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 208; SEQ ID NO: 209; and SEQ ID NO: 210 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 63 or the heavy chain sequence of SEQ ID NO: 64.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 201 encoding the light chain variable sequenceof SEQ ID NO: 61; the polynucleotide SEQ ID NO: 202 encoding the lightchain sequence of SEQ ID NO: 62; the polynucleotide SEQ ID NO: 203encoding the heavy chain variable sequence of SEQ ID NO: 63; thepolynucleotide SEQ ID NO: 204 encoding the heavy chain sequence of SEQID NO: 64; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 205; SEQ ID NO: 206; and SEQ ID NO: 207) of thelight chain variable sequence of SEQ ID NO: 61 or the light chainsequence of SEQ ID NO: 62; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 208; SEQ ID NO: 209; andSEQ ID NO: 210) of the heavy chain variable sequence of SEQ ID NO: 63 orthe heavy chain sequence of SEQ ID NO: 64.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab7, the polynucleotidesencoding the full length Ab7 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 202 encoding the light chain sequenceof SEQ ID NO: 62 and the polynucleotide SEQ ID NO: 204 encoding theheavy chain sequence of SEQ ID NO: 64.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab7 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab7 or Fab fragments thereofmay be produced via expression of Ab7 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab8

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 71:

(SEQ ID NO: 211) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTACAATTACAACTACCTTGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTAGTACTGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 72:

(SEQ ID NO: 212) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTACAATTACAACTACCTTGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTAGTACTGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 73:

(SEQ ID NO: 213) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGACCTCAGTAACCACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCGTTGGTATCAATGGTCGCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCTAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 74:

(SEQ ID NO: 214) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGACCTCAGTAACCACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCGTTGGTATCAATGGTCGCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCTAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA TGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 215; SEQ ID NO: 216; and SEQ ID NO: 217 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 71 or the light chain sequence of SEQ ID NO: 72.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 218; SEQ ID NO: 219; and SEQ ID NO: 220 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 73 or the heavy chain sequence of SEQ ID NO: 74.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 211 encoding the light chain variable sequenceof SEQ ID NO: 71; the polynucleotide SEQ ID NO: 212 encoding the lightchain sequence of SEQ ID NO: 72; the polynucleotide SEQ ID NO: 213encoding the heavy chain variable sequence of SEQ ID NO: 73; thepolynucleotide SEQ ID NO: 214 encoding the heavy chain sequence of SEQID NO: 74; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 215; SEQ ID NO: 216; and SEQ ID NO: 217) of thelight chain variable sequence of SEQ ID NO: 71 or the light chainsequence of SEQ ID NO: 72; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 218; SEQ ID NO: 219; andSEQ ID NO: 220) of the heavy chain variable sequence of SEQ ID NO: 73 orthe heavy chain sequence of SEQ ID NO: 74.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab8, the polynucleotidesencoding the full length Ab8 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 212 encoding the light chain sequenceof SEQ ID NO: 72 and the polynucleotide SEQ ID NO: 214 encoding theheavy chain sequence of SEQ ID NO: 74.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab8 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab8 or Fab fragments thereofmay be produced via expression of Ab8 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab9

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 81:

(SEQ ID NO: 221) CAAGTGCTGACCCAGACTCCATCCCCCGTGTCTGCAGCTGTGGGAAGCACAGTCACCATCAATTGCCAGGCCAGTCAGAATGTTTATAATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCAACTGATCTATTCTACGTCCACTCTGGCATCTGGGGTCTCATCGCGATTCAGAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCAGTTATGATTGTAGTCGTGGTGATTGTTTTGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 82:

(SEQ ID NO: 222) CAAGTGCTGACCCAGACTCCATCCCCCGTGTCTGCAGCTGTGGGAAGCACAGTCACCATCAATTGCCAGGCCAGTCAGAATGTTTATAATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCAACTGATCTATTCTACGTCCACTCTGGCATCTGGGGTCTCATCGCGATTCAGAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCAGTTATGATTGTAGTCGTGGTGATTGTTTTGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 83:

(SEQ ID NO: 223) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGAATCGGCCTCAGTAGCTACTACATGCAGTGGGTCCGCCAGTCTCCAGGGAGGGGGCTGGAATGGATCGGAGTCATTGGTAGTGATGGTAAGACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAAGACCTCGTCGACCACGGTGGATCTGAGAATGGCCAGTCTGACAACCGAGGACACGGCCACCTATTTCTGTACCAGAGGGGACATCTGGGGCCCGGGGACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 84:

(SEQ ID NO: 224) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGAATCGGCCTCAGTAGCTACTACATGCAGTGGGTCCGCCAGTCTCCAGGGAGGGGGCTGGAATGGATCGGAGTCATTGGTAGTGATGGTAAGACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAAGACCTCGTCGACCACGGTGGATCTGAGAATGGCCAGTCTGACAACCGAGGACACGGCCACCTATTTCTGTACCAGAGGGGACATCTGGGGCCCGGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 225; SEQ ID NO: 226; and SEQ ID NO: 227 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 81 or the light chain sequence of SEQ ID NO: 82.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 228; SEQ ID NO: 229; and SEQ ID NO: 230 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 83 or the heavy chain sequence of SEQ ID NO: 84.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 221 encoding the light chain variable sequenceof SEQ ID NO: 81; the polynucleotide SEQ ID NO: 222 encoding the lightchain sequence of SEQ ID NO: 82; the polynucleotide SEQ ID NO: 223encoding the heavy chain variable sequence of SEQ ID NO: 83; thepolynucleotide SEQ ID NO: 224 encoding the heavy chain sequence of SEQID NO: 84; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 225; SEQ ID NO: 226; and SEQ ID NO: 227) of thelight chain variable sequence of SEQ ID NO: 81 or the light chainsequence of SEQ ID NO: 82; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 228; SEQ ID NO: 229; andSEQ ID NO: 230) of the heavy chain variable sequence of SEQ ID NO: 83 orthe heavy chain sequence of SEQ ID NO: 84.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab9, the polynucleotidesencoding the full length Ab9 antibody comprise, or alternatively consistof, the polynucleotide SEQ ID NO: 222 encoding the light chain sequenceof SEQ ID NO: 82 and the polynucleotide SEQ ID NO: 224 encoding theheavy chain sequence of SEQ ID NO: 84.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab9 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab9 or Fab fragments thereofmay be produced via expression of Ab9 polynucleotides in mammalian cellssuch as CHO, NSO or HEK 293 cells, fungal, insect, or microbial systemssuch as yeast cells (for example diploid yeast such as diploid Pichia)and other yeast strains. Suitable Pichia species include, but are notlimited to, Pichia pastoris.

Antibody Ab10

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 91:

(SEQ ID NO: 231) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAATGTTTACAATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTAGTCGTGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO: 92:

(SEQ ID NO: 232) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAATGTTTACAATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTAGTCGTGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 93:

(SEQ ID NO: 233) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGGCCTCAGTAGCTACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTAGTGATGGTAAGACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTACCAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO: 94:

(SEQ ID NO: 234) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGGCCTCAGTAGCTACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTAGTGATGGTAAGACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTACCAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA TGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 235; SEQ ID NO: 236; and SEQ ID NO: 237 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 91 or the light chain sequence of SEQ ID NO: 92.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 238; SEQ ID NO: 239; and SEQ ID NO:240 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 93 or the heavy chain sequence of SEQ ID NO: 94.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 231 encoding the light chain variable sequenceof SEQ ID NO: 91; the polynucleotide SEQ ID NO: 232 encoding the lightchain sequence of SEQ ID NO: 92; the polynucleotide SEQ ID NO: 233encoding the heavy chain variable sequence of SEQ ID NO: 93; thepolynucleotide SEQ ID NO: 234 encoding the heavy chain sequence of SEQID NO: 94; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 235; SEQ ID NO: 236; and SEQ ID NO: 237) of thelight chain variable sequence of SEQ ID NO: 91 or the light chainsequence of SEQ ID NO: 92; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 238; SEQ ID NO: 239; andSEQ ID NO: 240) of the heavy chain variable sequence of SEQ ID NO: 93 orthe heavy chain sequence of SEQ ID NO: 94.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab10, the polynucleotidesencoding the full length Ab10 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 232 encoding the light chainsequence of SEQ ID NO: 92 and the polynucleotide SEQ ID NO: 234 encodingthe heavy chain sequence of SEQ ID NO: 94.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab10 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab10 or Fab fragmentsthereof may be produced via expression of Ab10 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab11

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 101:

(SEQ ID NO: 241) CAGGTGCTGACCCAGACTGCATCCCCCGTGTCTCCAGCTGTGGGAAGCACAGTCACCATCAATTGCCGGGCCAGTCAGAGTGTTTATTATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCAGTTATGATTGTAGTAATGGTGATTGTTTTGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:102:

(SEQ ID NO: 242) CAGGTGCTGACCCAGACTGCATCCCCCGTGTCTCCAGCTGTGGGAAGCACAGTCACCATCAATTGCCGGGCCAGTCAGAGTGTTTATTATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCAGTTATGATTGTAGTAATGGTGATTGTTTTGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTT AG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 103:

(SEQ ID NO: 243) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGAGGATCCCTGACACTCACCTGCACAGTCTCTGGAATCGACGTCACTAACTACTATATGCAATGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGTCATTGGTGTGAATGGTAAGAGATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGCGACATCTGGGGCCCGGGGACCCTCGTCACCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:104:

(SEQ ID NO: 244) CAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGAGGATCCCTGACACTCACCTGCACAGTCTCTGGAATCGACGTCACTAACTACTATATGCAATGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAGTCATTGGTGTGAATGGTAAGAGATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGCGACATCTGGGGCCCGGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAA ATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 245; SEQ ID NO: 246; and SEQ ID NO: 247 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 101 or the light chain sequence of SEQ ID NO: 102.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 248; SEQ ID NO: 249; and SEQ ID NO: 250 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 103 or the heavy chain sequence of SEQ ID NO: 104.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 241 encoding the light chain variable sequenceof SEQ ID NO: 101; the polynucleotide SEQ ID NO: 242 encoding the lightchain sequence of SEQ ID NO: 102; the polynucleotide SEQ ID NO: 243encoding the heavy chain variable sequence of SEQ ID NO: 103; thepolynucleotide SEQ ID NO: 244 encoding the heavy chain sequence of SEQID NO: 104; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 245; SEQ ID NO: 246; and SEQ ID NO: 247) of thelight chain variable sequence of SEQ ID NO: 101 or the light chainsequence of SEQ ID NO: 102; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 248; SEQ ID NO: 249; andSEQ ID NO: 250) of the heavy chain variable sequence of SEQ ID NO: 103or the heavy chain sequence of SEQ ID NO: 104.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab11, the polynucleotidesencoding the full length Ab11 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 242 encoding the light chainsequence of SEQ ID NO: 102 and the polynucleotide SEQ ID NO: 244encoding the heavy chain sequence of SEQ ID NO: 104.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab11 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab11 or Fab fragmentsthereof may be produced via expression of Ab11 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab12

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 111:

(SEQ ID NO: 251) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCGGGCCAGTCAGAGTGTTTACTATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTAGTAATGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAA CGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:112:

(SEQ ID NO: 252) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCGGGCCAGTCAGAGTGTTTACTATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTAGTAATGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 113:

(SEQ ID NO: 253) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGACGTCACTAACTACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTGTGAATGGTAAGAGATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCCAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTC GAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:114:

(SEQ ID NO: 254) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGACGTCACTAACTACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTGTGAATGGTAAGAGATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTGCCAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 255; SEQ ID NO: 256; and SEQ ID NO: 257 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 111 or the light chain sequence of SEQ ID NO: 112.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 258; SEQ ID NO: 259; and SEQ ID NO: 260 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 113 or the heavy chain sequence of SEQ ID NO: 114.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 251 encoding the light chain variable sequenceof SEQ ID NO: 111; the polynucleotide SEQ ID NO: 252 encoding the lightchain sequence of SEQ ID NO: 112; the polynucleotide SEQ ID NO: 253encoding the heavy chain variable sequence of SEQ ID NO: 113; thepolynucleotide SEQ ID NO: 254 encoding the heavy chain sequence of SEQID NO: 114; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 255; SEQ ID NO: 256; and SEQ ID NO: 257) of thelight chain variable sequence of SEQ ID NO: 111 or the light chainsequence of SEQ ID NO: 112; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 258; SEQ ID NO: 259; andSEQ ID NO: 260) of the heavy chain variable sequence of SEQ ID NO: 113or the heavy chain sequence of SEQ ID NO: 114.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab12, the polynucleotidesencoding the full length Ab12 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 252 encoding the light chainsequence of SEQ ID NO: 112 and the polynucleotide SEQ ID NO: 254encoding the heavy chain sequence of SEQ ID NO: 114.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab12 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab12 or Fab fragmentsthereof may be produced via expression of Ab12 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab13

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 121:

(SEQ ID NO: 261) GCCATCGTGATGACCCAGACTCCATCTTCCAAGTCTGTCCCTGTGGGAGACACAGTCACCATCAATTGCCAGGCCAGTGAGAGTCTTTATAATAACAACGCCTTGGCCTGGTTTCAGCAGAAACCAGGGCAGCCTCCCAAGCGCCTGATCTATGATGCATCCAAACTGGCATCTGGGGTCCCATCGCGGTTCAGTGGCGGTGGGTCTGGGACACAGTTCACTCTCACCATCAGTGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTGGAGGCTACAGAAGTGATAGTGTTGATGGTGTTGCTTTCGCCGGAGGGACCGAGGTGGTGGTCAAA CGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:122:

(SEQ ID NO: 262) GCCATCGTGATGACCCAGACTCCATCTTCCAAGTCTGTCCCTGTGGGAGACACAGTCACCATCAATTGCCAGGCCAGTGAGAGTCTTTATAATAACAACGCCTTGGCCTGGTTTCAGCAGAAACCAGGGCAGCCTCCCAAGCGCCTGATCTATGATGCATCCAAACTGGCATCTGGGGTCCCATCGCGGTTCAGTGGCGGTGGGTCTGGGACACAGTTCACTCTCACCATCAGTGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTGGAGGCTACAGAAGTGATAGTGTTGATGGTGTTGCTTTCGCCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 123:

(SEQ ID NO: 263) CAGTCGGTGGAGGAGTCCGGGGGAGGCCTGGTCCAGCCTGAGGGATCCCTGACACTCACCTGCACAGCCTCTGGATTCGACTTCAGTAGCAATGCAATGTGGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGATGCATTTACAATGGTGATGGCAGCACATACTACGCGAGCTGGGTGAATGGCCGATTCTCCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAACTGAATAGTCTGACAGTCGCGGACACGGCCACGTATTATTGTGCGAGAGATCTTGACTTGTGGGGCCCGGGCACCCTCGTCA CCGTCTCGAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:124:

(SEQ ID NO: 264) CAGTCGGTGGAGGAGTCCGGGGGAGGCCTGGTCCAGCCTGAGGGATCCCTGACACTCACCTGCACAGCCTCTGGATTCGACTTCAGTAGCAATGCAATGTGGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGATGCATTTACAATGGTGATGGCAGCACATACTACGCGAGCTGGGTGAATGGCCGATTCTCCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAACTGAATAGTCTGACAGTCGCGGACACGGCCACGTATTATTGTGCGAGAGATCTTGACTTGTGGGGCCCGGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 265; SEQ ID NO: 266; and SEQ ID NO: 267 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 121 or the light chain sequence of SEQ ID NO: 122.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 268; SEQ ID NO: 269; and SEQ ID NO: 270 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 123 or the heavy chain sequence of SEQ ID NO: 124.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 261 encoding the light chain variable sequenceof SEQ ID NO: 121; the polynucleotide SEQ ID NO: 262 encoding the lightchain sequence of SEQ ID NO: 122; the polynucleotide SEQ ID NO: 263encoding the heavy chain variable sequence of SEQ ID NO: 123; thepolynucleotide SEQ ID NO: 264 encoding the heavy chain sequence of SEQID NO: 124; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 265; SEQ ID NO: 266; and SEQ ID NO: 267) of thelight chain variable sequence of SEQ ID NO: 121 or the light chainsequence of SEQ ID NO: 122; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 268; SEQ ID NO: 269; andSEQ ID NO: 270) of the heavy chain variable sequence of SEQ ID NO: 123or the heavy chain sequence of SEQ ID NO: 124.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab13, the polynucleotidesencoding the full length Ab13 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 262 encoding the light chainsequence of SEQ ID NO: 122 and the polynucleotide SEQ ID NO: 264encoding the heavy chain sequence of SEQ ID NO: 124.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab13 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab13 or Fab fragmentsthereof may be produced via expression of Ab13 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

Antibody Ab14

The invention is further directed to polynucleotides encoding antibodypolypeptides having binding specificity to CGRP. In one embodiment ofthe invention, polynucleotides of the invention comprise, oralternatively consist of, the following polynucleotide sequence encodingthe variable light chain polypeptide sequence of SEQ ID NO: 131:

(SEQ ID NO: 271) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAATGTTTACAATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTAGTCGTGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAA CGT.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the light chain polypeptide sequence of SEQ ID NO:132:

(SEQ ID NO: 272) CAAGTGCTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAATGTTTACAATAACAACTACCTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCAACTGATCTATTCTACATCCACTCTGGCATCTGGGGTCCCATCTCGTTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCTGGGCAGTTATGATTGTAGTCGTGGTGATTGTTTTGTTTTCGGCGGAGGAACCAAGGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG.

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 133:

(SEQ ID NO: 273) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGGCCTCAGTAGCTACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTAGTGATGGTAAGACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTACCAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTC GAGC.

In one embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the heavy chain polypeptide sequence of SEQ ID NO:134:

(SEQ ID NO: 274) GAGGTGCAGCTTGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGTCTCTGGAATCGGCCTCAGTAGCTACTACATGCAATGGGTCCGTCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCGGAGTCATTGGTAGTGATGGTAAGACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAGAGACAATTCCAAGACCACGGTGTATCTTCAAATGAACAGCCTGAGAGCTGAGGACACTGCTGTGTATTTCTGTACCAGAGGGGACATCTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACGCGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 275; SEQ ID NO: 276; and SEQ ID NO: 277 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 131 or the light chain sequence of SEQ ID NO: 132.

In a further embodiment of the invention, polynucleotides encodingantibody fragments having binding specificity to CGRP comprise, oralternatively consist of, one or more of the polynucleotide sequences ofSEQ ID NO: 278; SEQ ID NO: 279; and SEQ ID NO: 280 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 133 or the heavy chain sequence of SEQ ID NO: 134.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding antibody fragments having binding specificity to CGRP comprise,or alternatively consist of, one, two, three or more, including all ofthe following polynucleotides encoding antibody fragments: thepolynucleotide SEQ ID NO: 271 encoding the light chain variable sequenceof SEQ ID NO: 131; the polynucleotide SEQ ID NO: 272 encoding the lightchain sequence of SEQ ID NO: 132; the polynucleotide SEQ ID NO: 273encoding the heavy chain variable sequence of SEQ ID NO: 133; thepolynucleotide SEQ ID NO: 274 encoding the heavy chain sequence of SEQID NO: 134; polynucleotides encoding the complementarity-determiningregions (SEQ ID NO: 275; SEQ ID NO: 276; and SEQ ID NO: 277) of thelight chain variable sequence of SEQ ID NO: 131 or the light chainsequence of SEQ ID NO: 132; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 278; SEQ ID NO: 279; andSEQ ID NO: 280) of the heavy chain variable sequence of SEQ ID NO: 133or the heavy chain sequence of SEQ ID NO: 134.

In a preferred embodiment of the invention, polynucleotides of theinvention comprise, or alternatively consist of, polynucleotidesencoding Fab (fragment antigen binding) fragments having bindingspecificity for CGRP. With respect to antibody Ab14, the polynucleotidesencoding the full length Ab14 antibody comprise, or alternativelyconsist of, the polynucleotide SEQ ID NO: 272 encoding the light chainsequence of SEQ ID NO: 132 and the polynucleotide SEQ ID NO: 274encoding the heavy chain sequence of SEQ ID NO: 134.

Another embodiment of the invention contemplates these polynucleotidesincorporated into an expression vector for expression in mammalian cellssuch as CHO, NSO, HEK-293, or in fungal, insect, or microbial systemssuch as yeast cells such as the yeast Pichia. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris. In one embodiment ofthe invention described herein (infra), Fab fragments may be produced byenzymatic digestion (e.g., papain) of Ab14 following expression of thefull-length polynucleotides in a suitable host. In another embodiment ofthe invention, anti-CGRP antibodies such as Ab14 or Fab fragmentsthereof may be produced via expression of Ab14 polynucleotides inmammalian cells such as CHO, NSO or HEK 293 cells, fungal, insect, ormicrobial systems such as yeast cells (for example diploid yeast such asdiploid Pichia) and other yeast strains. Suitable Pichia speciesinclude, but are not limited to, Pichia pastoris.

In one embodiment, the invention is directed to an isolatedpolynucleotide comprising a polynucleotide encoding an anti-CGRP V_(H)antibody amino acid sequence selected from SEQ ID NO: 3, 13, 23, 33, 43,53, 63, 73, 83, 93, 103, 113, 123, or 133, or encoding a variant thereofwherein at least one framework residue (FR residue) has been substitutedwith an amino acid present at the corresponding position in a rabbitanti-CGRP antibody V_(H) polypeptide or a conservative amino acidsubstitution.

In another embodiment, the invention is directed to an isolatedpolynucleotide comprising the polynucleotide sequence encoding ananti-CGRP V_(L) antibody amino acid sequence of 1, 11, 21, 31, 41, 51,61, 71, 81, 91, 101, 111, 121, or 131, or encoding a variant thereofwherein at least one framework residue (FR residue) has been substitutedwith an amino acid present at the corresponding position in a rabbitanti-CGRP antibody V_(L) polypeptide or a conservative amino acidsubstitution.

In yet another embodiment, the invention is directed to one or moreheterologous polynucleotides comprising a sequence encoding thepolypeptides contained in SEQ ID NO:1 and SEQ ID NO:3; SEQ ID NO:11 andSEQ ID NO:13; SEQ ID NO:21 and SEQ ID NO:23; SEQ ID NO:31 and SEQ IDNO:33; SEQ ID NO:41 and SEQ ID NO:43; SEQ ID NO:51 and SEQ ID NO:53, SEQID NO:61 and SEQ ID NO:63; SEQ ID NO:71 and SEQ ID NO:73; SEQ ID NO:81and SEQ ID NO:83; SEQ ID NO:91 and SEQ ID NO:93; SEQ ID NO:101 and SEQID NO:103; SEQ ID NO:111 and SEQ ID NO:113; SEQ ID NO:121 and SEQ IDNO:123; or SEQ ID NO:131 and SEQ ID NO:133.

In another embodiment, the invention is directed to an isolatedpolynucleotide that expresses a polypeptide containing at least one CDRpolypeptide derived from an anti-CGRP antibody wherein said expressedpolypeptide alone specifically binds CGRP or specifically binds CGRPwhen expressed in association with another polynucleotide sequence thatexpresses a polypeptide containing at least one CDR polypeptide derivedfrom an anti-CGRP antibody wherein said at least one CDR is selectedfrom those contained in the V_(L) or V_(H) polypeptides of SEQ ID NO: 1,3, 11, 13, 21, 23, 31, 33, 41, 43, 51, 53, 61, 63, 71, 73, 81, 83, 91,93, 101, 103, 111, 113, 121, 123, 131, or SEQ ID NO:133.

Host cells and vectors comprising said polynucleotides are alsocontemplated.

The invention further contemplates vectors comprising the polynucleotidesequences encoding the variable heavy and light chain polypeptidesequences, as well as the individual complementarity-determining regions(CDRs, or hypervariable regions), as set forth herein, as well as hostcells comprising said vector sequences. In one embodiment of theinvention, the host cell is a yeast cell. In another embodiment of theinvention, the yeast host cell belongs to the genus Pichia.

B-Cell Screening and Isolation

In one embodiment, the present invention contemplates the preparationand isolation of a clonal population of antigen-specific B cells thatmay be used for isolating at least one CGRP antigen-specific cell, whichcan be used to produce a monoclonal antibody against CGRP, which isspecific to a desired CGRP antigen, or a nucleic acid sequencecorresponding to such an antibody. Methods of preparing and isolatingsaid clonal population of antigen-specific B cells are taught, forexample, in U.S. patent publication no. US 2007/0269868 toCarvalho-Jensen et al., the disclosure of which is herein incorporatedby reference in its entirety. Methods of preparing and isolating saidclonal population of antigen-specific B cells are also taught herein inthe examples. Methods of “enriching” a cell population by size ordensity are known in the art. See, e.g., U.S. Pat. No. 5,627,052. Thesesteps can be used in addition to enriching the cell population byantigen-specificity.

Methods of Humanizing Antibodies

In another embodiment, the present invention contemplates methods forhumanizing antibody heavy and light chains. Methods for humanizingantibody heavy and light chains which may be applied to anti-CGRPantibodies are taught, for example, in U.S. patent applicationpublication no. US 2009/0022659 to Olson et al., and in U.S. Pat. No.7,935,340 to Garcia-Martinez et al., the disclosures of each of whichare herein incorporated by reference in their entireties.

Methods of Producing Antibodies and Fragments Thereof

In another embodiment, the present invention contemplates methods forproducing anti-CGRP antibodies and fragments thereof. Methods forproducing anti-CGRP antibodies and fragments thereof secreted frompolyploidal, preferably diploid or tetraploid strains of matingcompetent yeast are taught, for example, in U.S. patent applicationpublication no. US 2009/0022659 to Olson et al., and in U.S. Pat. No.7,935,340 to Garcia-Martinez et al., the disclosures of each of whichare herein incorporated by reference in their entireties.

Other methods of producing antibodies are well known to those ofordinary skill in the art. For example, methods of producing chimericantibodies are now well known in the art (See, for example, U.S. Pat.No. 4,816,567 to Cabilly et al.; Morrison et al., P.N.A.S. USA,81:8651-55 (1984); Neuberger, M. S. et al., Nature, 314:268-270 (1985);Boulianne, G. L. et al., Nature, 312:643-46 (1984), the disclosures ofeach of which are herein incorporated by reference in their entireties).

Likewise, other methods of producing humanized antibodies are now wellknown in the art (See, for example, U.S. Pat. Nos. 5,530,101, 5,585,089,5,693,762, and 6,180,370 to Queen et al; U.S. Pat. Nos. 5,225,539 and6,548,640 to Winter; U.S. Pat. Nos. 6,054,297, 6,407,213 and 6,639,055to Carter et al; U.S. Pat. No. 6,632,927 to Adair; Jones, P. T. et al,Nature, 321:522-525 (1986); Reichmann, L., et al, Nature, 332:323-327(1988); Verhoeyen, M, et al, Science, 239:1534-36 (1988), thedisclosures of each of which are herein incorporated by reference intheir entireties).

Antibody polypeptides of the invention having CGRP binding specificitymay also be produced by constructing, using conventional techniques wellknown to those of ordinary skill in the art, an expression vectorcontaining an operon and a DNA sequence encoding an antibody heavy chainin which the DNA sequence encoding the CDRs required for antibodyspecificity is derived from a non-human cell source, preferably a rabbitB-cell source, while the DNA sequence encoding the remaining parts ofthe antibody chain is derived from a human cell source.

A second expression vector is produced using the same conventional meanswell known to those of ordinary skill in the art, said expression vectorcontaining an operon and a DNA sequence encoding an antibody light chainin which the DNA sequence encoding the CDRs required for antibodyspecificity is derived from a non-human cell source, preferably a rabbitB-cell source, while the DNA sequence encoding the remaining parts ofthe antibody chain is derived from a human cell source.

The expression vectors are transfected into a host cell by conventiontechniques well known to those of ordinary skill in the art to produce atransfected host cell, said transfected host cell cultured byconventional techniques well known to those of ordinary skill in the artto produce said antibody polypeptides.

The host cell may be co-transfected with the two expression vectorsdescribed above, the first expression vector containing DNA encoding anoperon and a light chain-derived polypeptide and the second vectorcontaining DNA encoding an operon and a heavy chain-derived polypeptide.The two vectors contain different selectable markers, but preferablyachieve substantially equal expression of the heavy and light chainpolypeptides. Alternatively, a single vector may be used, the vectorincluding DNA encoding both the heavy and light chain polypeptides. Thecoding sequences for the heavy and light chains may comprise cDNA,genomic DNA, or both.

The host cells used to express the antibody polypeptides may be either abacterial cell such as E. coli, or a eukaryotic cell such as P.pastoris. In one embodiment of the invention, a mammalian cell of awell-defined type for this purpose, such as a myeloma cell, a Chinesehamster ovary (CHO) cell line, a NSO cell line, or a HEK293 cell linemay be used.

The general methods by which the vectors may be constructed,transfection methods required to produce the host cell and culturingmethods required to produce the antibody polypeptides from said hostcells all include conventional techniques. Although preferably the cellline used to produce the antibody is a mammalian cell line, any othersuitable cell line, such as a bacterial cell line such as an E.coli-derived bacterial strain, or a yeast cell line, may alternativelybe used.

Similarly, once produced the antibody polypeptides may be purifiedaccording to standard procedures in the art, such as for examplecross-flow filtration, ammonium sulphate precipitation, affinity columnchromatography and the like.

The antibody polypeptides described herein may also be used for thedesign and synthesis of either peptide or non-peptide mimetics thatwould be useful for the same therapeutic applications as the antibodypolypeptides of the invention. See, for example, Saragobi et al,Science, 253:792-795 (1991), the contents of which is hereinincorporated by reference in its entirety.

Screening Assays

The invention also includes screening assays designed to assist in theidentification of diseases and disorders associated with CGRP inpatients exhibiting symptoms of a CGRP associated disease or disorder.

In one embodiment of the invention, the anti-CGRP antibodies of theinvention, or CGRP binding fragments thereof, are used to detect thepresence of CGRP in a biological sample obtained from a patientexhibiting symptoms of a disease or disorder associated with CGRP. Thepresence of CGRP, or elevated levels thereof when compared topre-disease levels of CGRP in a comparable biological sample, may bebeneficial in diagnosing a disease or disorder associated with CGRP.

Another embodiment of the invention provides a diagnostic or screeningassay to assist in diagnosis of diseases or disorders associated withCGRP in patients exhibiting symptoms of a CGRP associated disease ordisorder identified herein, comprising assaying the level of CGRPexpression in a biological sample from said patient using apost-translationally modified anti-CGRP antibody or binding fragmentthereof. The anti-CGRP antibody or binding fragment thereof may bepost-translationally modified to include a detectable moiety such as setforth previously in the disclosure.

The CGRP level in the biological sample is determined using a modifiedanti-CGRP antibody or binding fragment thereof as set forth herein, andcomparing the level of CGRP in the biological sample against a standardlevel of CGRP (e.g., the level in normal biological samples). Theskilled clinician would understand that some variability may existbetween normal biological samples, and would take that intoconsideration when evaluating results. In one embodiment of theinvention, the anti-CGRP antibodies of the invention may be used tocorrelate CGRP expression levels with a particular stage of cancerousdevelopment. One skilled in the art would be able to measure CGRP innumerous subjects in order to establish ranges of CGRP expression thatcorrespond to clinically defined stages of cancerous development. Theseranges will allow the skilled practitioner to measure CGRP in a subjectdiagnosed with a cancer and correlate the levels in each subject with arange that corresponds to a stage of said cancer. One skilled in the artwould understand that by measuring CGRP in the patient at differentintervals, the progression of the cancer can be determined.

The above-recited assay may also be useful in monitoring a disease ordisorder, where the level of CGRP obtained in a biological sample from apatient believed to have a CGRP associated disease or disorder iscompared with the level of CGRP in prior biological samples from thesame patient, in order to ascertain whether the CGRP level in saidpatient has changed with, for example, a treatment regimen.

The invention is also directed to a method of in vivo imaging whichdetects the presence of cells which express CGRP comprisingadministering a diagnostically effective amount of a diagnosticcomposition. Said in vivo imaging is useful for the detection or imagingof CGRP expressing tumors or metastases, for example, and can be usefulas part of a planning regimen for the design of an effective cancertreatment protocol. The treatment protocol may include, for example, oneor more of radiation, chemotherapy, cytokine therapy, gene therapy, andantibody therapy, as well as an anti-CGRP antibody or fragment thereof.

The present invention further provides for a kit for detecting bindingof an anti-CGRP antibody of the invention to CGRP. In particular, thekit may be used to detect the presence of a CGRP specifically reactivewith an anti-CGRP antibody of the invention or an immunoreactivefragment thereof. The kit may also include an antibody bound to asubstrate, a secondary antibody reactive with the antigen and a reagentfor detecting a reaction of the secondary antibody with the antigen.Such a kit may be an ELISA kit and can comprise the substrate, primaryand secondary antibodies when appropriate, and any other necessaryreagents such as detectable moieties, enzyme substrates, and colorreagents, for example as described herein. The diagnostic kit may alsobe in the form of an immunoblot kit. The diagnostic kit may also be inthe form of a chemiluminescent kit (Meso Scale Discovery, Gaithersburg,Md.). The diagnostic kit may also be a lanthanide-based detection kit(PerkinElmer, San Jose, Calif.).

A skilled clinician would understand that a biological sample includes,but is not limited to, sera, plasma, urine, saliva, mucous, pleuralfluid, synovial fluid and spinal fluid.

Methods of Ameliorating or Reducing Symptoms of or Treating, orPreventing, Diseases and Disorders Associated with, CGRP

In another embodiment of the invention, anti-CGRP antibodies describedherein, or fragments thereof, are useful for ameliorating or reducingthe symptoms of, or treating, or preventing, diseases and disordersassociated with CGRP. Anti-CGRP antibodies described herein, orfragments thereof, as well as combinations, can also be administered ina therapeutically effective amount to patients in need of treatment ofdiseases and disorders associated with CGRP in the form of apharmaceutical composition as described in greater detail below.

In another embodiment of the invention, anti-CGRP antibodies describedherein, or fragments thereof, are useful for ameliorating or reducingthe symptoms of, or treating, or preventing, migraines (with or withoutaura), weight loss, cancer or tumors, angiogenesis associated withcancer or tumor growth, angiogenesis associated with cancer or tumorsurvival, pain, hemiplagic migraines, cluster headaches, migrainousneuralgia, chronic headaches, tension headaches, general headaches, hotflushes, chronic paroxysomal hemicrania, secondary headaches due to anunderlying structural problem in the head or neck, cranial neuralgia,sinus headaches (such as for example associated with sinusitis), andallergy-induced headaches or migraines.

In one embodiment of the invention, anti-CGRP antibodies describedherein, or fragments thereof and/or with a second agent, are useful forameliorating or reducing the symptoms of, or treating, or preventing,the following non-limiting listing of diseases and disorders: pain,inflammatory pain, post-operative incision pain, complex regional painsyndrome, cancer pain, primary or metastatic bone cancer pain, fracturepain, chronic pain, osteoporotic fracture pain, pain resulting fromburn, osteoporosis, gout joint pain, abdominal pain, pain associatedwith sickle cell crises, and other nociceptic pain, as well ashepatocellular carcinoma, breast cancer, liver cirrhosis, neurogenicpain, neuropathic pain, nociceptic pain, trigeminal neuralgia,post-herpetic neuralgia, phantom limb pain, fibromyalgia, menstrualpain, ovarialgia, reflex sympathetic dystrophy, neurogenic pain,osteoarthritis or rheumatoid arthritis pain, lower back pain, diabeticneuropathy, sciatica, or pain or visceral pain associated with:gastro-esophageal reflux, dyspepsia, irritable bowel syndrome, irritablecolon, spastic colon, mucous colitis, inflammatory bowel disease,Crohn's disease, ileitis, ulcerative colitis, renal colic, dysmenorrhea,cystitis, menstrual period, labor, menopause, prostatitis, pancreatitis,renal colic, dysmenorrhea, cystitis, including interstitial cystitis(IC), surgery associated with the ileus, diverticulitis, peritonitis,pericarditis, hepatitis, appendicitis, colitis, cholecystitis,endometriosis, chronic and/or acute pancreatitis, myocardial infarction,kidney pain, pleural pain, prostatitis, pelvic pain, trauma to an organ,chronic nociceptive pain, chronic neuropathic pain, chronic inflammatorypain, fibromyalgia, breakthrough pain and persistent pain, and cancerpain arising from malignancy or from cancer preferably selected from oneor more of: adenocarcinoma in glandular tissue, blastoma in embryonictissue of organs, carcinoma in epithelial tissue, leukemia in tissuesthat form blood cells, lymphoma in lymphatic tissue, myeloma in bonemarrow, sarcoma in connective or supportive tissue, adrenal cancer,AIDS-related lymphoma, anemia, bladder cancer, bone cancer, braincancer, breast cancer, carcinoid tumours, cervical cancer, chemotherapy,colon cancer, cytopenia, endometrial cancer, esophageal cancer, gastriccancer, head cancer, neck cancer, hepatobiliary cancer, kidney cancer,leukemia, liver cancer, lung cancer, lymphoma, Hodgkin's disease,lymphoma, non-Hodgkin's, nervous system tumours, oral cancer, ovariancancer, pancreatic cancer, prostate cancer, rectal cancer, skin cancer,stomach cancer, testicular cancer, thyroid cancer, urethral cancer, bonecancer, sarcomas cancer of the connective tissue, cancer of bone tissue,cancer of blood-forming cells, cancer of bone marrow, multiple myeloma,leukaemia, primary or secondary bone cancer, tumours that metastasize tothe bone, tumours infiltrating the nerve and hollow viscus, tumours nearneural structures. Further preferably the cancer pain comprises visceralpain, preferably visceral pain which arises from pancreatic cancerand/or metastases in the abdomen. Further preferably the cancer paincomprises somatic pain, preferably somatic pain due to one or more ofbone cancer, metastasis in the bone, postsurgical pain, sarcomas cancerof the connective tissue, cancer of bone tissue, cancer of blood-formingcells of the bone marrow, multiple myeloma, leukaemia, primary orsecondary bone cancer.

In another embodiment of the invention, anti-CGRP antibodies describedherein, or fragments thereof and/or with a second agent, are useful forameliorating or reducing the symptoms of, or treating, or preventing,the following non-limiting listing of diseases and disorders: cancer ortumors, angiogenesis associated with cancer or tumor growth,angiogenesis associated with cancer or tumor survival.

In another embodiment of the invention, anti-CGRP antibodies describedherein, or fragments thereof and/or with a second agent, are useful forameliorating or reducing the symptoms of, or treating, or preventing,the following non-limiting listing of diseases and disorders:neurogenic, neuropathic or nociceptic pain. Neuropathic pain mayinclude, but is not limited to, trigeminal neuralgia, post-herpeticneuralgia, phantom limb pain, fibromyalgia, menstrual pain, ovarialgia,reflex sympathetic dystrophy and neurogenic pain. In other preferredembodiments, osteoarthritis or rheumatoid arthritis pain, lower backpain, diabetic neuropathy, sciatica, and other neuropathic pain.

In another embodiment of the invention, anti-CGRP antibodies describedherein, or fragments thereof and/or with a second agent, are useful forameliorating or reducing the symptoms of, or treating, or preventing,the following non-limiting listing of diseases and disorders: overactivebladder and other urinary conditions, gastro-esophageal reflux andvisceral pain associated with gastro-esophageal reflux, dyspepsia,irritable bowel syndrome, inflammatory bowel disease, Crohn's disease,ileitis, ulcerative colitis, renal colic, dysmenorrhea, cystitis,menstrual period, labor, menopause, prostatitis, pruritis, orpancreatitis. Also, the subject CGRP antibodies and antibody fragmentsmay be used alone or in conjunction with other active agents, e.g.,opioids and non-opioid analgesics such as NSAIDs to elicit analgesia orto potentiate the efficacy of another analgesic or to prevent oralleviate tolerance to a specific analgesic such as morphine or relatedopioid analgesics. Evidence for role of CGRP in blocking/reversingdevelopment of morphine—induced analgesia: is the fact that CGRP8-37 andCGRP Receptor antagonist (BIBN4096BS) reportedly prevent/reversedevelopment of morphine tolerance—(Powell et al., 2000 J Brit JPharmacol (131):875; Menard et al., 1996 J Neurosci (16):2342; Wang etal., 2009 FASEB J (23):2576; Wang et al., 2010 Pain (151):194)

The subject antibodies potentially may be combined with any opioidanalgesic or NSAID or other analgesic, potentially another antibody, inorder to increase or enhance pain management, or to reverse or suppresstolerance to an analgesic such as an opioid analgesic compound. This mayallow for such analgesic compounds to be administered for longerduration or at reduced dosages thereby potentially alleviating adverseside effects associated therewith.

The term “opioid analgesic” herein refers to all drugs, natural orsynthetic, with morphine-like actions. The synthetic and semi-syntheticopioid analgesics are derivatives of five chemical classes of compound:phenanthrenes; phenylheptylamines; phenylpiperidines; morphinans; andbenzomorphans, all of which are within the scope of the term. Exemplaryopioid analgesics include codeine, dihydrocodeine, diacetylmorphine,hydrocodone, hydromorphone, levorphanol, oxymorphone, alfentanil,buprenorphine, butorphanol, fentanyl, sufentanyl, meperidine, methadone,nalbuphine, propoxyphene and pentazocine or pharmaceutically acceptablesalts thereof.

The term “NSAID” refers to a non-steroidal anti-inflammatory compound.NSAIDs are categorized by virtue of their ability to inhibitcyclooxygenase. Cyclooxygenase 1 and cyclooxygenase 2 are two majorisoforms of cyclooxygenase and most standard NSAIDs are mixed inhibitorsof the two isoforms. Most standard NSAIDs fall within one of thefollowing five structural categories: (1) propionic acid derivatives,such as ibuprofen, naproxen, naprosyn, diclofenac, and ketoprofen; (2)acetic acid derivatives, such as tolmetin and slindac; (3) fenamic acidderivatives, such as mefenamic acid and meclofenamic acid; (4)biphenylcarboxylic acid derivatives, such as diflunisal and flufenisal;and (5) oxicams, such as piroxim, sudoxicam, and isoxicam. Another classof NSAID has been described which selectively inhibit cyclooxygenase 2.Cox-2 inhibitors have been described, e.g., in U.S. Pat. Nos. 5,616,601;5,604,260; 5,593,994; 5,550,142; 5,536,752; 5,521,213; 5,475,995;5,639,780; 5,604,253; 5,552,422; 5,510,368; 5,436,265; 5,409,944; and5,130,311, all of which are hereby incorporated by reference. Certainexemplary COX-2 inhibitors include celecoxib (SC-58635), DUP-697,flosulide (CGP-28238), meloxicam, 6-methoxy-2 naphthylacetic acid(6-MNA), rofecoxib, MK-966, nabumetone (prodrug for 6-MNA), nimesulide,NS-398, SC-5766, SC-58215, T-614; or combinations thereof.

In some embodiments, aspirin and/or acetaminophen may be taken inconjunction with the subject CGRP antibody or fragment. Aspirin isanother type of non-steroidal anti-inflammatory compound.

Exemplary, non-limiting diseases and disorders that can be treatedand/or prevented by the administration of the CGRP antibodies of thepresent invention include, pain resulting from any condition associatedwith neurogenic, neuropathic, inflammatory, thermal or nociceptic pain.Preferably the disorder will be associated with increased CGRP at thepain site. In certain embodiments of neuropathic pain, referredtrigeminal neuralgia, post-herpetic neuralgia, phantom limb pain,fibromyalgia, reflex sympathetic dystrophy and neurogenic painconditions are preferably treated. In other embodiments, cancer pain,particularly, bone cancer pain, osteoarthritis or rheumatoid arthritispain, lower back pain, post-operative incision pain, fracture pain,osteoporotic fracture pain, osteoporosis, gout joint pain, diabeticneuropathy, sciatica, pains associated with sickle cell crises,migraine, and other neuropathic and/or nociceptic pain are preferablytreated. Thus, the present invention includes methods of treating,preventing, and/or ameliorating any disease or disorder associated withCGRP activity or CGRP upregulation (including any of the above mentionedexemplary diseases, disorders and conditions) through use of theantibodies and antibody fragments of the invention. The therapeuticmethods of the present invention comprise administering to a subject anyformulation comprising an anti-CGRP antibody as disclosed herein aloneor in association with another active agent.

The subject to which the pharmaceutical formulation is administered canbe, e.g., any human or non-human animal that is in need of suchtreatment, prevention and/or amelioration, or who would otherwisebenefit from the inhibition or attenuation of CGRP-mediated activity.For example, the subject can be an individual that is diagnosed with, orwho is deemed to be at risk of being afflicted by any of theaforementioned diseases or disorders. The present invention furtherincludes the use of any of the pharmaceutical formulations disclosedherein in the manufacture of a medicament for the treatment, preventionand/or amelioration of any disease or disorder associated with CGRPactivity (including any of the above mentioned exemplary diseases,disorders and conditions).

Administration

In one embodiment of the invention, the anti-CGRP antibodies describedherein, or CGRP binding fragments thereof, as well as combinations ofsaid antibodies or antibody fragments, are administered to a subject ata concentration of between about 0.1 and 100.0 mg/kg of body weight ofrecipient subject. In a preferred embodiment of the invention, theanti-CGRP antibodies described herein, or CGRP binding fragmentsthereof, as well as combinations of said antibodies or antibodyfragments, are administered to a subject at a concentration of about 0.4mg/kg of body weight of recipient subject. In a preferred embodiment ofthe invention, the anti-CGRP antibodies described herein, or CGRPbinding fragments thereof, as well as combinations of said antibodies orantibody fragments, are administered to a recipient subject with afrequency of once every twenty-six weeks or less, such as once everysixteen weeks or less, once every eight weeks or less, once every fourweeks or less, once every two weeks or less, once every week or less, oronce daily or less.

Fab fragments may be administered every two weeks or less, every week orless, once daily or less, multiple times per day, and/or every fewhours. In one embodiment of the invention, a patient receives Fabfragments of 0.1 mg/kg to 40 mg/kg per day given in divided doses of 1to 6 times a day, or in a sustained release form, effective to obtaindesired results.

It is to be understood that the concentration of the antibody or Fabadministered to a given patient may be greater or lower than theexemplary administration concentrations set forth above in paragraphs[0552] and [0553].

A person of skill in the art would be able to determine an effectivedosage and frequency of administration through routine experimentation,for example guided by the disclosure herein and the teachings inGoodman, L. S., Gilman, A., Brunton, L. L., Lazo, J. S., & Parker, K. L.(2006). Goodman & Gilman's the pharmacological basis of therapeutics.New York: McGraw-Hill; Howland, R. D., Mycek, M. J., Harvey, R. A.,Champe, P. C., & Mycek, M. J. (2006). Pharmacology. Lippincott'sillustrated reviews. Philadelphia: Lippincott Williams & Wilkins; andGolan, D. E. (2008). Principles of pharmacology: the pathophysiologicbasis of drug therapy. Philadelphia, Pa., [etc.]: Lippincott Williams &Wilkins.

In another embodiment of the invention, the anti-CGRP antibodiesdescribed herein, or CGRP binding fragments thereof, as well ascombinations of said antibodies or antibody fragments, are administeredto a subject in a pharmaceutical formulation.

A “pharmaceutical composition” refers to a chemical or biologicalcomposition suitable for administration to a mammal. Such compositionsmay be specifically formulated for administration via one or more of anumber of routes, including but not limited to buccal, epicutaneous,epidural, inhalation, intraarterial, intracardial,intracerebroventricular, intradermal, intramuscular, intranasal,intraocular, intraperitoneal, intraspinal, intrathecal, intravenous,oral, parenteral, rectally via an enema or suppository, subcutaneous,subdermal, sublingual, transdermal, and transmucosal. In addition,administration can occur by means of injection, powder, liquid, gel,drops, or other means of administration.

In one embodiment of the invention, the anti-CGRP antibodies describedherein, or CGRP binding fragments thereof, as well as combinations ofsaid antibodies or antibody fragments, may be optionally administered incombination with one or more active agents. Such active agents includeanalgesic, anti-histamine, antipyretic, anti-inflammatory, antibiotic,antiviral, and anti-cytokine agents. Active agents include agonists,antagonists, and modulators of TNF-α, IL-2, IL-4, IL-6, IL-10, IL-12,IL-13, IL-18, IFN-α, IFN-γ, BAFF, CXCL13, IP-10, VEGF, EPO, EGF, HRG,Hepatocyte Growth Factor (HGF), Hepcidin, including antibodies reactiveagainst any of the foregoing, and antibodies reactive against any oftheir receptors. Active agents also include but are not limited to2-Arylpropionic acids, Aceclofenac, Acemetacin, Acetylsalicylic acid(Aspirin), Alclofenac, Alminoprofen, Amoxiprin, Ampyrone, Arylalkanoicacids, Azapropazone, Benorylate/Benorilate, Benoxaprofen, Bromfenac,Carprofen, Celecoxib, Choline magnesium salicylate, Clofezone, COX-2inhibitors, Dexibuprofen, Dexketoprofen, Diclofenac, Diflunisal,Droxicam, Ethenzamide, Etodolac, Etoricoxib, Faislamine, fenamic acids,Fenbufen, Fenoprofen, Flufenamic acid, Flunoxaprofen, Flurbiprofen,Ibuprofen, Ibuproxam, Indometacin, Indoprofen, Kebuzone, Ketoprofen,Ketorolac, Lornoxicam, Loxoprofen, Lumiracoxib, Magnesium salicylate,Meclofenamic acid, Mefenamic acid, Meloxicam, Metamizole, Methylsalicylate, Mofebutazone, Nabumetone, Naproxen, N-Arylanthranilic acids,Nerve Growth Factor (NGF), Oxametacin, Oxaprozin, Oxicams,Oxyphenbutazone, Parecoxib, Phenazone, Phenylbutazone, Phenylbutazone,Piroxicam, Pirprofen, profens, Proglumetacin, Pyrazolidine derivatives,Rofecoxib, Salicyl salicylate, Salicylamide, Salicylates, Substance P,Sulfinpyrazone, Sulindac, Suprofen, Tenoxicam, Tiaprofenic acid,Tolfenamic acid, Tolmetin, and Valdecoxib.

An anti-histamine can be any compound that opposes the action ofhistamine or its release from cells (e.g., mast cells). Anti-histaminesinclude but are not limited to acrivastine, astemizole, azatadine,azelastine, betatastine, brompheniramine, buclizine, cetirizine,cetirizine analogues, chlorpheniramine, clemastine, CS 560,cyproheptadine, desloratadine, dexchlorpheniramine, ebastine,epinastine, fexofenadine, HSR 609, hydroxyzine, levocabastine,loratidine, methscopolamine, mizolastine, norastemizole, phenindamine,promethazine, pyrilamine, terfenadine, and tranilast.

Antibiotics include but are not limited to Amikacin, Aminoglycosides,Amoxicillin, Ampicillin, Ansamycins, Arsphenamine, Azithromycin,Azlocillin, Aztreonam, Bacitracin, Carbacephem, Carbapenems,Carbenicillin, Cefaclor, Cefadroxil, Cefalexin, Cefalothin, Cefalotin,Cefamandole, Cefazolin, Cefdinir, Cefditoren, Cefepime, Cefixime,Cefoperazone, Cefotaxime, Cefoxitin, Cefpodoxime, Cefprozil,Ceftazidime, Ceftibuten, Ceftizoxime, Ceftobiprole, Ceftriaxone,Cefuroxime, Cephalosporins, Chloramphenicol, Cilastatin, Ciprofloxacin,Clarithromycin, Clindamycin, Cloxacillin, Colistin, Co-trimoxazole,Dalfopristin, Demeclocycline, Dicloxacillin, Dirithromycin, Doripenem,Doxycycline, Enoxacin, Ertapenem, Erythromycin, Ethambutol,Flucloxacillin, Fosfomycin, Furazolidone, Fusidic acid, Gatifloxacin,Geldanamycin, Gentamicin, Glycopeptides, Herbimycin, Imipenem,Isoniazid, Kanamycin, Levofloxacin, Lincomycin, Linezolid, Lomefloxacin,Loracarbef, Macrolides, Mafenide, Meropenem, Meticillin, Metronidazole,Mezlocillin, Minocycline, Monobactams, Moxifloxacin, Mupirocin,Nafcillin, Neomycin, Netilmicin, Nitrofurantoin, Norfloxacin, Ofloxacin,Oxacillin, Oxytetracycline, Paromomycin, Penicillin, Penicillins,Piperacillin, Platensimycin, Polymyxin B, Polypeptides, Prontosil,Pyrazinamide, Quinolones, Quinupristin, Rifampicin, Rifampin,Roxithromycin, Spectinomycin, Streptomycin, Sulfacetamide,Sulfamethizole, Sulfanilimide, Sulfasalazine, Sulfisoxazole,Sulfonamides, Teicoplanin, Telithromycin, Tetracycline, Tetracyclines,Ticarcillin, Timidazole, Tobramycin, Trimethoprim,Trimethoprim-Sulfamethoxazole, Troleandomycin, Trovafloxacin, andVancomycin.

Active agents also include Aldosterone, Beclometasone, Betamethasone,Corticosteroids, Cortisol, Cortisone acetate, Deoxycorticosteroneacetate, Dexamethasone, Fludrocortisone acetate, Glucocorticoids,Hydrocortisone, Methylprednisolone, Prednisolone, Prednisone, Steroids,and Triamcinolone. Any suitable combination of these active agents isalso contemplated.

A “pharmaceutical excipient” or a “pharmaceutically acceptableexcipient” is a carrier, usually a liquid, in which an activetherapeutic agent is formulated. In one embodiment of the invention, theactive therapeutic agent is a humanized antibody described herein, orone or more fragments thereof. The excipient generally does not provideany pharmacological activity to the formulation, though it may providechemical and/or biological stability, and release characteristics.Exemplary formulations can be found, for example, in Remington'sPharmaceutical Sciences, 19^(th) Ed., Grennaro, A., Ed., 1995 which isincorporated by reference.

As used herein “pharmaceutically acceptable carrier” or “excipient”includes any and all solvents, dispersion media, coatings, antibacterialand antifungal agents, isotonic and absorption delaying agents that arephysiologically compatible. In one embodiment, the carrier is suitablefor parenteral administration. Alternatively, the carrier can besuitable for intravenous, intraperitoneal, intramuscular, or sublingualadministration. Pharmaceutically acceptable carriers include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. The use of such media and agents for pharmaceuticallyactive substances is well known in the art. Except insofar as anyconventional media or agent is incompatible with the active compound,use thereof in the pharmaceutical compositions of the invention iscontemplated. Supplementary active compounds can also be incorporatedinto the compositions.

Pharmaceutical compositions typically must be sterile and stable underthe conditions of manufacture and storage. The invention contemplatesthat the pharmaceutical composition is present in lyophilized form. Thecomposition can be formulated as a solution, microemulsion, liposome, orother ordered structure suitable to high drug concentration. The carriercan be a solvent or dispersion medium containing, for example, water,ethanol, polyol (for example, glycerol, propylene glycol, and liquidpolyethylene glycol), and suitable mixtures thereof. The inventionfurther contemplates the inclusion of a stabilizer in the pharmaceuticalcomposition. The proper fluidity can be maintained, for example, by themaintenance of the required particle size in the case of dispersion andby the use of surfactants.

In many cases, it will be preferable to include isotonic agents, forexample, sugars, polyalcohols such as mannitol, sorbitol, or sodiumchloride in the composition. Prolonged absorption of the injectablecompositions can be brought about by including in the composition anagent which delays absorption, for example, monostearate salts andgelatin. Moreover, the alkaline polypeptide can be formulated in a timerelease formulation, for example in a composition which includes a slowrelease polymer. The active compounds can be prepared with carriers thatwill protect the compound against rapid release, such as a controlledrelease formulation, including implants and microencapsulated deliverysystems. Biodegradable, biocompatible polymers can be used, such asethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters, polylactic acid and polylactic, polyglycolic copolymers(PLG). Many methods for the preparation of such formulations are knownto those skilled in the art.

For each of the recited embodiments, the compounds can be administeredby a variety of dosage forms. Any biologically-acceptable dosage formknown to persons of ordinary skill in the art, and combinations thereof,are contemplated. Examples of such dosage forms include, withoutlimitation, reconstitutable powders, elixirs, liquids, solutions,suspensions, emulsions, powders, granules, particles, microparticles,dispersible granules, cachets, inhalants, aerosol inhalants, patches,particle inhalants, implants, depot implants, injectables (includingsubcutaneous, intramuscular, intravenous, and intradermal), infusions,and combinations thereof.

The above description of various illustrated embodiments of theinvention is not intended to be exhaustive or to limit the invention tothe precise form disclosed. While specific embodiments of, and examplesfor, the invention are described herein for illustrative purposes,various equivalent modifications are possible within the scope of theinvention, as those skilled in the relevant art will recognize. Theteachings provided herein of the invention can be applied to otherpurposes, other than the examples described above.

These and other changes can be made to the invention in light of theabove detailed description. In general, in the following claims, theterms used should not be construed to limit the invention to thespecific embodiments disclosed in the specification and the claims.Accordingly, the invention is not limited by the disclosure, but insteadthe scope of the invention is to be determined entirely by the followingclaims.

The invention may be practiced in ways other than those particularlydescribed in the foregoing description and examples. Numerousmodifications and variations of the invention are possible in light ofthe above teachings and, therefore, are within the scope of the appendedclaims.

Certain teachings related to methods for obtaining a clonal populationof antigen-specific B cells were disclosed in U.S. Provisional patentapplication No. 60/801,412, filed May 19, 2006, the disclosure of whichis herein incorporated by reference in its entirety.

Certain teachings related to humanization of rabbit-derived monoclonalantibodies and preferred sequence modifications to maintain antigenbinding affinity were disclosed in International Application No.PCT/US2008/064421, corresponding to International Publication No.WO/2008/144757, entitled “Novel Rabbit Antibody Humanization Methods andHumanized Rabbit Antibodies”, filed May 21, 2008, the disclosure ofwhich is herein incorporated by reference in its entirety.

Certain teachings related to producing antibodies or fragments thereofusing mating competent yeast and corresponding methods were disclosed inU.S. patent application Ser. No. 11/429,053, filed May 8, 2006, (U.S.Patent Application Publication No. US2006/0270045), the disclosure ofwhich is herein incorporated by reference in its entirety.

Certain CGRP antibody polynucleotides and polypeptides are disclosed inthe sequence listing accompanying this patent application filing, andthe disclosure of said sequence listing is herein incorporated byreference in its entirety.

The entire disclosure of each document cited (including patents, patentapplications, journal articles, abstracts, manuals, books, or otherdisclosures) in the Background of the Invention, Detailed Description,and Examples is herein incorporated by reference in their entireties.

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the subject invention, and are not intended to limit thescope of what is regarded as the invention. Efforts have been made toensure accuracy with respect to the numbers used (e.g. amounts,temperature, concentrations, etc.) but some experimental errors anddeviations should be allowed for. Unless otherwise indicated, parts areparts by weight, molecular weight is average molecular weight,temperature is in degrees centigrade; and pressure is at or nearatmospheric.

EXAMPLES Example 1 Preparation of Antibodies that Bind CGRP

By using the antibody selection protocol described herein, one cangenerate an extensive panel of antibodies.

Immunization Strategy

Rabbits were immunized with human CGRPα (American Peptides, SunnyvaleCalif. and Bachem, Torrance Calif.). Immunization consisted of a firstsubcutaneous (sc) injection of 100 μg of antigen mixed with 100 μg ofKLH in complete Freund's adjuvant (CFA) (Sigma) followed by two boosts,two weeks apart each containing 50 μg antigen mixed with 50 μg inincomplete Freund's adjuvant (IFA) (Sigma). Animals were bled on day 55,and serum titers were determined by ELISA (antigen recognition) and byinhibition of CGRP driven cAMP increase in SK-N-MC.

Antibody Selection Titer Assessment

To identify and characterize antibodies that bind to human CGRPcL,antibody-containing solutions were tested by ELISA. Briefly, neutravidincoated plates (Thermo Scientific), were coated with N-term biotinylatedhuman CGRPα (50 μL per well, 1 μg/mL) diluted in ELISA buffer (0.5% fishskin gelatin in PBS pH 7.4,) either for approximately 1 hr at roomtemperature or alternatively overnight at 4° C. The plates were thenfurther blocked with ELISA buffer for one hour at room temperature andwashed using wash buffer (PBS, 0.05% tween 20). Serum samples testedwere serially diluted using ELISA buffer. Fifty microliters of dilutedserum samples were transferred onto the wells and incubated for one hourat room temperature for one hour. After this incubation, the plate waswashed with wash buffer. For development, an anti-rabbit specificFc-HARP (1:5000 dilution in ELISA buffer) was added onto the wells andincubated for 45 min at RT. After a 3× wash step with wash solution, theplate was developed using TMB substrate for two minutes at roomtemperature and the reaction was quenched using 0.5M HCl. The wellabsorbance was read at 450 nm.

Titer Determination of Serum Samples by Functional Activity (Inhibitionof CGRP Driven cAMP Levels)

To identify and characterize antibodies with functional activity, aninhibition of CGRP driven increase of cAMP levels assay was done usingelectrochemiluminescence (Meso Scale Discovery, MSD). Briefly, antibodypreparations to be tested were serially diluted in MSD assay buffer(Hepes, MgC12, pH 7.3, 1 mg/mL blocker A, Meso Scale Discovery) in a 96well round bottom polystyrene plate (Costar). To this plate, human CGRPαwas added (10 ng/mL final concentration) diluted in MSD assay buffer andincubated for one hour at 37 C. Appropriate controls were used assuggested by the assay-kit manufacturer. Human neuroepithelioma cells(SK-N-MC, ATCC) were detached using an EDTA solution (5 mM in PBS) andwashed using growth media (MEM, 10% FBS, antibiotics) by centrifugation.The cell number was adjusted to 2 million cells per mL in assay buffer,and IBMX (3-Isobutyl-1-Methylxanthine, Sigma) was added to a finalconcentration of 0.2 mM right before loading cells onto cAMP assayplate. After the antibody human CGRPα solution was incubated for onehour 20 microliters of solution containing cells were transferred to thecAMP assay plate. All tested samples were run in duplicates withappropriate controls. Ten microliters of cells were added to the wellsand the plate was incubated for 30 minutes with shaking at roomtemperature. While cells were being incubated with the CGRP solution,the stop solution was prepared by making a 1:200 solution of TAG labeledcAMP (MSD) in lysis buffer (MSD). To stop the cells-CGRP incubation, 20microliters of stop solution was added to the cells and the plate wasincubated for one hour with shaking at room temperature. The read buffer(MSD) was diluted four times with water and 100 microliters were addedto all wells on the plate. The plate was then read using a Sector Imager2400 (MSD) and the Prism software was used for data fit and IC50determination.

Tissue Harvesting

Once acceptable titers were established, the rabbit(s) were sacrificed.Spleen, lymph nodes, and whole blood were harvested and processed asfollows:

Spleen and lymph nodes were processed into a single cell suspension bydisassociating the tissue and pushing through sterile wire mesh at 70 μm(Fisher) with a plunger of a 20 cc syringe. Cells were collected in PBS.Cells were washed twice by centrifugation. After the last wash, celldensity was determined by trypan blue. Cells were centrifuged at 1500rpm for 10 minutes; the supernatant was discarded. Cells wereresuspended in the appropriate volume of 10% dimethyl sulfoxide (DMSO,Sigma) in FBS (Hyclone) and dispensed at 1 ml/vial. Vials were stored at−70° C. in a slow freezing chamber for 24 hours and stored in liquidnitrogen.

Peripheral blood mononuclear cells (PBMCs) were isolated by mixing wholeblood with equal parts of the low glucose medium described above withoutFBS. 35 ml of the whole blood mixture was carefully layered onto 8 ml ofLympholyte Rabbit (Cedarlane) into a 45 ml conical tube (Corning) andcentrifuged 30 minutes at 2500 rpm at room temperature without brakes.After centrifugation, the PBMC layers were carefully removed using aglass Pasteur pipette (VWR), combined, and placed into a clean 50 mlvial. Cells were washed twice with the modified medium described aboveby centrifugation at 1500 rpm for 10 minutes at room temperature, andcell density was determined by trypan blue staining. After the lastwash, cells were resuspended in an appropriate volume of 10% DMSO/FBSmedium and frozen as described above.

B Cell Selection, Enrichment and Culture Conditions

On the day of setting up B cell culture, PBMC, splenocyte, or lymph nodevials were thawed for use. Vials were removed from LN2 tank and placedin a 37° C. water bath until thawed. Contents of vials were transferredinto 15 ml conical centrifuge tube (Corning) and 10 ml of modified RPMIdescribed above was slowly added to the tube. Cells were centrifuged for5 minutes at 2K RPM, and the supernatant was discarded. Cells wereresuspended in 10 ml of fresh media. Cell density and viability wasdetermined by trypan blue.

a) The Following Protocol was Used for Ab1 and Ab13

Cells were pre-mixed with the biotinylated human CGRPα as follows. Cellswere washed again and resuspended at 1E07 cells/80 μL medium.Biotinylated human CGRPα was added to the cell suspension at the finalconcentration of 5 ug/mL and incubated for 30 minutes at 4° C. Unboundbiotinylated human CGRPα was removed performing two 10 ml washes usingPBF [Ca/Mg free PBS (Hyclone), 2 mM ethylenediamine tetraacetic acid(EDTA), 0.5% bovine serum albumin (BSA) (Sigma-biotin free)]. After thesecond wash, cells were resuspended at 1E07 cells/80 μl PBF and 20 μl ofMACS® streptavidin beads (Miltenyi Biotech, Auburn Calif.) per 10E7cells were added to the cell suspension. Cells and beads were incubatedat 4° C. for 15 minutes and washed once with 2 ml of PBF per 10E7 cells.

b) The Following Protocol was Used for Ab4, Ab7, Ab9 and Ab11:

Biotinylated human CGRPα was pre-loaded onto the streptavidin beads asfollows. Seventy five microliters of streptavidin beads (Milteny Biotec,Auburn Calif.) were mixed with N-terminally biotinylated huCGRPα (10ug/ml final concentration) and 300 μl PBF. This mixture was incubated at4° C. for 30 min and unbound biotinylated human CGRPα was removed usinga MACS® separation column (Miltenyi Biotec, with a 1 ml rinse to removeunbound material. Then material was plunged out, then used to resuspendcells from above in 100u1 per 1E7 cells, the mixture was then incubatedat 4° C. for 30 min and washed once with 10 ml of PBF.

For both a) and b) protocols the following applied: After washing, thecells were resuspended in 500 μl of PBF and set aside. A MACS® MS column(Miltenyi Biotec, Auburn Calif.) was pre-rinsed with 500 ml of PBF on amagnetic stand (Milteni). Cell suspension was applied to the columnthrough a pre-filter, and unbound fraction was collected. The column waswashed with 2.5 ml of PBF buffer. The column was removed from the magnetstand and placed onto a clean, sterile 1.5 ml eppendorf tube. 1 ml ofPBF buffer was added to the top of the column, and positive selectedcells were collected. The yield and viability of positive cell fractionwas determined by trypan blue staining. Positive selection yielded anaverage of 1% of the starting cell concentration.

A pilot cell screen was established to provide information on seedinglevels for the culture. Plates were seeded at 10, 25, 50, 100, or 200enriched B cells/well. In addition, each well contained 50K cells/wellof irradiated EL-4.B5 cells (5,000 Rads) and an appropriate level ofactivated rabbit T cell supernatant (See U.S. Patent ApplicationPublication No. 20070269868)(ranging from 1-5% depending on preparation)in high glucose modified RPMI medium at a final volume of 250 μl/well.Cultures were incubated for 5 to 7 days at 37° C. in 4% CO₂.

B-Cell Culture Screening by Antigen-Recognition (ELISA)

To identify wells producing anti-human CGRPα antibodies, the sameprotocol as described for titer determination of serum samples byantigen-recognition (ELISA) was used with the following changes.Briefly, neutravidin coated plates were coated with a mixture of both N-and C-terminally biotinylated human CGRPα (50 μL per well, 1 μg/mLeach). B-cell supernatant samples (50 μL) were tested without priordilution.

Identification of Functional Activity in B-Cell Supernatants Using CGRPDriven Camp Production

To determine functional activity contained in B-cell supernatants, asimilar procedure to that described for the determination of functionaltiter of serum samples was used with the following modifications.Briefly, B-cell supernatant (20 μL) were used in place of the dilutedpolyclonal serum samples.

Isolation of Antigen-Specific B-Cells

Plates containing wells of interest were removed from −70° C., and thecells from each well were recovered using five washes of 200 microlitersof medium (10% RPMI complete, 55 μM BME) per well. The recovered cellswere pelleted by centrifugation and the supernatant was carefullyremoved. Pelleted cells were resuspended in 100 μl of medium. Toidentify antibody expressing cells, streptavidin coated magnetic beads(M280 dynabeads, Invitrogen) were coated with a combination of both N-and C-terminal biotinylated human CGRPα. Individual biotinylated humanCGRPα lots were optimized by serial dilution. One hundred microliterscontaining approximately 4×10E7 coated beads were then mixed with theresuspended cells. To this mixture 15 microliters of goat anti-rabbitH&L IgG-FITC (Jackson Immunoresearch) diluted 1:100 in medium wereadded.

Twenty microliters of cell/beads/anti-rabbit H&L suspension were removedand microliter droplets were dispensed on a one-well glass slidepreviously treated with Sigmacote (Sigma) totaling 35 to 40 droplets perslide. An impermeable barrier of paraffin oil (JT Baker) was used tosubmerge the droplets, and the slide was incubated for 90 minutes at 37°C. in a 4% CO2 incubator in the dark.

Specific B cells that produce antibody can be identified by thefluorescent ring around produced by the antibody secretion, recognitionof the bead-associated biotinylated antigen, and subsequent detection bythe fluorescent-IgG detection reagent. Once a cell of interest wasidentified it was recovered via a micromanipulator (Eppendorf). Thesingle cell synthesizing and exporting the antibody was transferred intoa microcentrifuge tube, frozen using dry ice and stored at −70° C.

Amplification and Sequence Determination of Antibody Sequences fromAntigen-Specific B Cells

Antibody sequences were recovered using a combined RT-PCR based methodfrom a single isolated B-cell. Primers containing restriction enzymeswere designed to anneal in conserved and constant regions of the targetimmunoglobulin genes (heavy and light), such as rabbit immunoglobulinsequences, and a two-step nested PCR recovery was used to amplify theantibody sequence. Amplicons from each well were analyzed for recoveryand size integrity. The resulting fragments are then digested with Alulto fingerprint the sequence clonality. Identical sequences displayed acommon fragmentation pattern in their electrophoretic analysis. Theoriginal heavy and light chain amplicon fragments were then digestedusing the restriction enzyme sites contained within the PCR primers andcloned into an expression vector. Vector containing subcloned DNAfragments were amplified and purified. Sequence of the subcloned heavyand light chains were verified prior to expression.

Recombinant Production of Monoclonal Antibody of Desired AntigenSpecificity and/or Functional Properties

To determine antigen specificity and functional properties of recoveredantibodies from specific B-cells, vectors driving the expression of thedesired paired heavy and light chain sequences were transfected intoHEK-293 cells.

Antigen-Recognition of Recombinant Antibodies by ELISA

To characterize recombinant expressed antibodies for their ability tobind to human-CGRPα, antibody-containing solutions were tested by ELISA.All incubations were done at room temperature. Briefly, Immulon IVplagtes (Thermo Scientific) were coated with a CGRPα containing solution(lut/mL in PBS) for 2 hours. CGRPα-coated plates were then washed threetimes in wash buffer (PBS, 0.05% Tween-20). The plates were then blockedusing a blocking solution (PBS, 0.5% fish skin gelatin, 0.05% Tween-20)for approximately one hour. The blocking solution was then removed andthe plates were then incubated with a dilution series of the antibodybeing tested for approximately one hour. At the end of this incubation,the plate was washed three times with wash buffer and further incubatedwith a secondary antibody containing solution (Peroxidase conjugatedaffinipure F(ab′)2 fragment goat anti-human IgG, Fc fragment specific(Jackson Immunoresearch) for approximately 45 minutes and washed threetimes. At that point a substrate solution (TMB peroxidase substrate,BioFx) and incubated for 3 to 5 minutes in the dark. The reaction wasstopped by addition of a HCl containing solution (0.5M) and the platewas read at 450 nm in a plate-reader.

Results: FIGS. 15-18 demonstrate that anti-CGRP antibodies Ab1-Ab14 bindto and recognize CGRPα.

Functional Characterization of Recombinant Antibodies by Modulation ofCGRP Driven Intracellular cAMP Levels and Cross Reactivity to Rats

To characterize recombinant expressed antibody for their ability toinhibit CGRPα mediated increased cellular levels of cAMP assay, anelectrochemiluminescence assay-kit (Meso Scale Discovery, MSD) was used.Briefly, antibody preparations to be tested were serially diluted in MSDassay buffer (Hepes, MgC12, pH 7.3, 1 mg/mL blocker A,Meso ScaleDiscovery) in a 96 well round bottom polystyrene plate (Costar). To thisplate, human CGRPcL was added (25 ng/mL final concentration) diluted inMSD assay buffer and incubated for one hour at 37° C. Appropriatecontrols were used as suggested by the assay-kit manufacturer. Humanneuroepithelioma cells (SK-N-MC, ATCC) were detached using an EDTAsolution (5 mM) and washed using growth media (MEM, 10% FBS,antibiotics) by centrifugation. The cell number was adjusted to 2million cells per mL in assay buffer, and IBMX(3-Isobutyl-1-Methylxanthine, 50 mM Sigma) was added to a finalconcentration of 0.2 mM right before loading cells onto cAMP assayplate. The antibody human CGRPα solution was incubated for one hourafter which 20 microliters of solution containing cells were transferredto the cAMP assay plate. All tested samples were run in duplicates withappropriate controls. Ten microliters of cells were added to the wellsand the plate was incubated for 30 minutes with shaking. While cellswere being incubated with the CGRP solution, the stop solution wasprepared by making a 1:200 solution of TAG labeled cAMP (MSD) in lysisbuffer (MSD). To stop the cells-CGRP incubation, 20 microliters of stopsolution was added to the cells and the plate was incubated for one hourwith shaking. The read buffer (MSD) was diluted four times with waterand 100 microliters were added to all wells on the plate. The plate wasthen read using a Sector Imager 2400 (MSD) and the Prism software wasused for data fit and IC50 determination.

To test for the ability of recombinant antibodies to antagonize humanCGRPβ a similar assay was performed with the substitution of the CGRPagonist (CGRPβ 10 ng/mL final concentration). Evaluation of therecombinant antibodies to recognize and inhibit rat CGRP-mediated cAMPgeneration was conducted using rat CGRP (5 ng/mL final concentration)and the rat L6 cell line (ATCC).

Results: FIGS. 19-37 demonstrate that anti-CGRP antibodies Ab1-Ab14inhibit CGRPα, CGRPβ, and rat CGRP mediated increased cellular levels ofcAMP.

Example 2 Enzymatic Production of Fab Fragments

Papain digestions were conducted using immobilized papain(Thermo/Pierce) as per manufacturer's instructions. Briefly, purifiedantibodies were incubated in a cystein/HC1-containing buffer withimmobilized papain at 37° C. with gentle rocking. The digestion wasmonitored by taking an aliquot and analyzing using SDS-PAGE for cleavageof the heavy chain. To stop the reaction, the immobilized papain wasspun out and washed using 50 mM Tris pH 7.5 and filtered. Undigestedfull length antibody and Fc fragments were removed by using aMabSelectSure (GE) column.

Example 3 Yeast Cell Expression

Construction of Pichia pastoris Expression Vectors for Heavy and LightChain.

The humanized light and heavy chain fragments were commerciallysynthesized and subcloned into a pGAP expression vector. The pGAPexpression vector uses the GAP promoter to drive expression of theimmunoglobulin chain and the human serum albumin (HSA) leader sequencefor export. In addition, this vector contains common elements such as abacterial origin of replication, and a copy of the kanamycin resistancegene which confers resistance to the antibiotic G418 in P. pastoris.G418 provides a means of selection for strains that contain the desiredexpression vector integrated into their genome.

Transformation of Expression Vectors into Haploid met1 and lys3 HostStrains of Pichia pastoris

All methods used for transformation of haploid P. pastoris strains andmanipulation of the P. pastoris sexual cycle were done as described inPichia Protocols (Methods in Molecular Biology Higgings, D R, and Cregg,J M, Eds. 1998. Humana Press, Totowa, N.J.). Prior to transformationeach vector was linearized within the GAP promoter sequences to directthe integration of the vector into the GAP promoter locus of the P.pastoris genome. Haploid strains were transfected using electroporationand successful transformants were selected on MPDS (yeast extract,peptone dextrose with sorbitol) G418 agar plates. Copy numbers of heavyand light chain genes were determined for haploid strains by Southernblot analysis. Haploid strains were then mated and selected for theirability to grow in the absence of the amino acid markers (i.e., Lys andMet). Resulting diploid clones were then subjected to a final Southernblot to confirm copy numbers of heavy and light chain genes. A cloneexpressing the antibody of interest was selected using biolayerinterferometry Protein-A biosensors to monitor expression (Octet,ForteBio).

Example 4 Expression of Ab3, Ab6 and Ab14 in Pichia pastoris

Three Pichia strains for expression of full-length antibody were made.For all the full length antibody expressing strains, haploids strainswere created and subsequently mated. One haploid strain expressedfull-length light chain sequence and another haploid strain expressedthe full-length heavy chain sequence. Each diploid strain was used togenerate a research cell bank and used for expression in a bioreactor.

First an inoculum was expanded using the research cell bank using mediumcomprised of the following nutrients (% w/v): yeast extract 3%,anhydrous dextrose 4%, YNB 1.34%, Biotin 0.004% and 100 mM potassiumphosphate. To generate the inoculum for the fermenters, the cell bankwas expanded for approximately 24 hours in a shaking incubator at 30° C.and 300 rpm. A 10% inoculum was then added to Labfors 2.5L workingvolume vessels containing 1 L sterile growth medium. The growth mediumwas comprised of the following nutrients: potassium sulfate 18.2 g/L,ammonium phosphate monobasic 36.4 g/L, potassium phosphate dibasic 12.8g/L, magnesium sulfate heptahydrate 3.72 g/L, sodium citrate dihydrate10 g/L, glycerol 40 g/L, yeast extract 30 g/L, PTM1 trace metals 4.35mL/L, and antifoam 204 1.67 mL/L. The PTM1 trace metal solution wascomprised of the following components: cupric sulfate pentahydrate 6g/L, sodium iodide 0.08 g/L, manganese sulfate hydrate 3 g/L, sodiummolybdate dihyrate 0.2 g/L, boric acid 0.02 g/L, cobalt chloride 0.5g/L, zinc chloride 20 g/L, ferrous sulfate heptahydrate 65 g/L, biotin0.2 g/L, and sulfuric acid 5 mL/L.

The bioreactor process control parameters were set as follows: Agitation1000 rpm, airflow 1.35 standard liter per minute, temperature 28° C. andpH was controlled at six using ammonium hydroxide. No oxygensupplementation was provided.

Fermentation cultures were grown for approximately 12 to 16 hours untilthe initial glycerol was consumed as denoted by a dissolved oxygenspike. The cultures were starved for approximately three hours after thedissolved oxygen spike. After this starvation period, a bolus additionof ethanol was added to the reactor to reach 1% ethanol (w/v). Thefermentation cultures were allowed to equilibrate for 15 to 30 minutes.Feed addition was initiated 30 minutes post-ethanol bolus and set at aconstant rate of 1 mL/min for 40 minutes, then the feed pump wascontrolled by an ethanol sensor keeping the concentration of ethanol at1% for the remainder of the run using an ethanol sensing probe (RavenBiotech). The feed was comprised of the following components: yeastextract 50 g/L, dextrose 500 g/L, magnesium sulfate heptahydrate 3 g/L,and PTM1 trace metals 12 mL/L. For fermentation of the full length Ab6and Ab14, sodium citrate dihydrate (0.5 g/L) was also added to the feed.The total fermentation time was approximately 90 hours.

Example 5 Methods of Humanizing Antibodies

Methods of humanizing antibodies have been described previously inissued U.S. Pat. No. 7,935,340, the disclosure of which is incorporatedherein by reference in its entirety. In some instances, a determinationof whether additional rabbit framework residues are required to maintainactivity is necessary. In some instances the humanized antibodies stillrequires some critical rabbit framework residues to be retained tominimize loss of affinity or activity. In these cases, it is necessaryto change single or multiple framework amino acids from human germlinesequences back to the original rabbit amino acids in order to havedesired activity. These changes are determined experimentally toidentify which rabbit residues are necessary to preserve affinity andactivity. This is now the end of the variable heavy and light chainhumanized amino acid sequence.

Example 6 Inhibition of CGRP Binding to its Cellular Receptor

To characterize recombinantly expressed antibodies for their ability toinhibit CGRP binding to its cellular receptor, a radioligand-bindingassay was performed as previously described [Elshourbagy et al,Endocrinology 139:1678 (1998); Zimmerman et al, Peptides, 16:421(1995)]. Membrane preparations of recombinant human CGRP receptors,calcitonin receptor-like receptor and RAMP1 (Chemiscreen, Millipore)were used. Antibody dilutions were preincubated with ¹²⁵I radiolabeldhuman CGRPα (0.03 nM) for 30 minutes at room temperature. Non-specificbinding was estimated in the presence of 0.1 μM human CGRPα. Membraneswere filtered and washed. The filters were then counted to determine¹²⁵I radiolabeld human CGRPα specifically bound.

Results: FIG. 38 demonstrates that anti-CGRP antibodies Ab1-Ab13 inhibitCGRP binding to its cellular receptor.

Example 7 Inhibition of Neurogenic Vasodilation by Anti-Cgrp Antibodiesin Rats

CGRP is a potent vasodilator (Nature 313: 54-56 (1985) and Br J. Clin.Pharmacol. 26(6):691-5. (1988)). A pharmacodynamic assay to measure CGRPreceptor antagonist activity non-invasively was used to characterizeanti-CGRP antibodies. The model relied on changes in dermal blood flowmeasured using a laser Doppler imaging following the topical applicationof a capsaicin solution. Capsaicin activates the transient receptorpotential vanilloid type 1 receptor (TRPV-1), producing neurogenicinflammation and vasodilatation via the local release of vasoactivemediators including CGRP and substance P (Br. J. Pharmacol. 110: 772-776(1993)).

On the day prior to the vasodilatation assay, animals were dosed withthe test agent or control via IP (intraperitoneal). Following dosing,the animals were shaved and depilated in the lower back region of theirdorsal side, in an area approximately 2×6 cm. The animals were thenreturned to their cages overnight. On the day of test, approximately 24hours post dosing, animals were anesthetized with isoflurane gas andplaced on a temperature controlled heating pad and fitted with a nosecone for continuous delivery of isoflurane. A laser Doppler imager wasused for the observation of vasodilatation. A beam of coherent red lightgenerated by a 633 nm helium-neon laser was directed to the shaved area,a rectangle (2×6 cm), and scanned at a medium resolution mode. Abaseline Doppler scan was obtained first and the location of O-ringplacement predetermined by identifying two similar low flux areas. Tworubber O-rings (˜1 cm in diameter) were placed in the selected regionsand a baseline scan was performed. Immediately after completion of thescan, 1 mg of capsaicin in 5 μL of an ethanol:acetone solution (1:1) wasapplied within each of the two O-rings Doppler scans were repeated at2.5, 5, 7.5, 10, 12.5, 15, 17.5, 20, 22.5, 25, 27.5 and 30 minutes afterthe application of capsaicin. Percent change from baseline mean Fluxwithin each of the two O-rings, was plotted as the results ofvasodilatation due to capsaicin.

In order to test recombinantly expressed antibodies for their ability toinhibit CGRP binding to its cellular receptor, a radioligand-bindingassay was performed as previously described.

Results: FIGS. 39 and 40 demonstrates that anti-CGRP antibodies Ab3 andAb6 reduced vasodilation in this model following capsaicinadministration.

Example 8 Effect of CGRP Antibody Administration on Overactive Bladder

Experiments were conducted to assess the potential efficacy of ananti-CGRP antibody administration on bladder continence and overactivebladder. Bladder continence is a balance between urethral closure anddetrusor muscle activity, and overactive bladder is a conditioncharacterized by urgency, urinary incontinence, frequency and nocturia.Some anecdotal evidence reported in the literature suggests that CGRPmay be involved with bladder continence and may correlate to and perhapsplay a causal role in overactive bladder disease pathology. Accordingly,it was hoped that the inventive anti-CGRP antibodies, especially giventheir high affinity to CGRP, would potentially help prevent or alleviatethis sometimes debilitating condition. (The evidence that that CGRP mayplay a role in overactive bladder includes the fact that CGRP is presentin the urinary tract, DRG and spinal cord (Wharton et al., 1986 Neurosci(3):727) Also, C-fiber afferents are critical for carrying impulsesinvolved in micturition to spinal cord (Yoshida et al., 2011 J PharmacolSci (112):128) and these fibers are affected by CGRP. Further, it hasbeen reported that intravesical administration of Botox suppresses CGRPand significantly reduces intercontraction interval in an acetic acidinduced bladder pain model (Chuang et al., 2004 J Urol (172):1529;Chuang et al., 2009 J Urol (182):786)). Moreover, it has been recentlyreported that the administration of an anti-CGRP antibody purportedlyreduces the number of bladder contractions in turpentine-oil—inducedoveractive bladder model (Pfizer PCT Patent Application WO2011/024113)).

Materials and Methods

Animals:

Female Sprague-Dawley rats (247-299 g) (Charles River Laboratories,Saint Germain sur l'Arbresle, France) were delivered to the laboratoryat least 5 days before the experiments in order to be acclimatized tolaboratory conditions. They were housed 3 per cage (polypropylene type Ecages size: 1032 cm²) and given food (Teklad 2016 global rodents,Harlan, 03800 Gannat, France) and water ad libitum. Sawdust (Souralit2912 plus, Souralit, 17080 Girona, Spain) bedding for rodent cages waschanged twice weekly. The animal room temperature (20±2° C.) wasmaintained with a 12/12 hour alternating light-dark cycle (light phase 7am: 7 pm) and relative humidity maintained at 40-70%.

Laboratory Equipment

Bladder catheters were connected via a T-tube to a strain gauge MX 860Novatrans III Gold (Medex Medical SARL, Nantes-Carquefou, France) and asyringe pump (70-2208 Model II plus, Harvard Apparatus, Les Ullis,France and Razel R-99E, Fisher Bioblock, Illkirch, France). Intravesicalpressure was recorded continuously using a PowerLab interface(ADInstruments Pty Ltd, Castle-Hill, Australia) and Chart® softwarerunning on a PC. Data were analyzed with Microsoft Excel® software.

Test Substances

Test Anti-CGRP antibody (Ab3)

Negative control antibody (Anti-digitoxin antibody).

Chemical Reagents

Physiological saline (NaCl 0.9%) (batch no 11043411, CAS no 7647-14-5)was purchased from B-Braun via Centravet (Lapalisse, France).

Anesthetic Substances

Urethane (batch no BCBC9294, CAS no 51-79-6) and sodium pentobarbital(batch no 150A1, CAS no 76-74-4) were supplied by Sigma-Aldrich (StQuentin Fallavier, France) and Centravet (Lapalisse, France),respectively.

Experimental Groups

Two experimental groups of 10 rats were used in the experiments. Eachgroup was administered 10 mg/kg of either the control or the anti-CGRPantibody:

Study Design

Experimental Procedure

Female rats were administered test antibody or negative control antibodyintravenously at a dose of 10 mg/kg, 18 hours prior to experiments usinga tail vein injection. Fifteen (15) hours later, rats were anesthetizedwith urethane (1.2 g/kg, subcutaneous (s.c.). Three (3) hours after thes.c. administration of urethane, a polyethylene catheter (0.58 and 0.96mm of internal and outer diameters, respectively) was inserted into thebladder through the dome and secured with a purse-string suture. Bodytemperature was maintained at 37±2° C. (TCAT-2LV controller, Physitemp,ADInstruments Pty Ltd., Casttle Hill, Australia) throughout theexperiment.

Cystometric Experiment

Cystometric investigations were performed in anesthetized female ratsafter surgery. Physiological saline at room temperature was continuouslyinfused into the bladder at a constant flow rate (2 mL/h) for a periodof at least 30 min.

At the end of the cystometric experiments, animals were sacrificed by alethal injection (1 mL) of sodium pentobarbital (54.7 mg/mL) (CAS no76-74-4) followed by cervical dislocation.

Cystometric Parameters

The cystometric parameters measured were:

Amplitude of micturition (AM), i.e. pressure between threshold pressureand Maximal pressure of micturition (mmHg),

Intercontraction interval (ICI), i.e. time between two subsequentmicturitions (sec),

Micturition frequency (MF), i.e. number of micturition contractions/15min (peaks/15 min).

Exclusion Criteria

Two rats were excluded during the experiments: One was excluded as itpresented with bladder hyperactivity during the saline intravesicalinfusion, and the other because the depth of anesthesia changed duringthe experiment inducing modifications of the cystometric profile.

Analysis of Results

For each rat, values for AM and ICI were calculated as the mean of thelast four or five micturitions during saline infusion. Values for MFwere calculated as the mean of micturitions obtained for two intervalsof 15 minutes during saline infusion.

Results are presented as mean values±standard error of the mean (±sem).Figures and statistical analyses were performed using GraphPad Prism®(Version 4; GraphPad Software Inc., La Jolla, Calif., USA).

Statistical comparisons of values (saline infusion) in the anti-CGRPantibody group versus the control antibody group were performed usingunpaired Student t-test.

A p<0.05 was accepted for statistical significance.

Results:

As shown in FIG. 41, ICI was significantly greater and MF wassignificantly lower in the anti-CGRP Ab-treated group (FIGS. 41A and Brespectively; p<0.05, unpaired Student t-test). No significantdifference was observed for AM between groups (FIG. 41C, p>0.05,unpaired Student t-test).

These results suggest that anti-CGRP antibodies may be useful inpreventing or alleviating overactive bladder, improving urinarycontinency and treatment of related urinary conditions.

Example 9 Relief of Neuropathic Pain in Rats

Damage to the peripheral nerves often leads to chronic referred painthat is neuropathic in origin. This pain syndrome consists ofsensitivity to external stimuli (e.g., mechanical and/or thermal) thatare not normally noxious. Consequently, neuropathic pain is refractoryto traditional analgesic approaches, making it difficult to treat.Experimentally, neuropathic pain can be modeled in animals via surgicaltrauma to peripheral nerves. The Chung model is one such system whereneuropathic pain is induced by ligation of the spinal nerves of L5 andL6.

In this Example, a spinal nerve ligation was performed on male SpragueDawley rats. They were tested for pain sensitivity on Day 13 (allodyniaconfirmation) and then again after each administration of Ab2 using thevon Frey test of mechanical allodynia to assess possible anti-allodynicactivity.

Methods

Male Sprague Dawley rats (Harlan Laboratories) weighing 200-225 g atarrival, were unpacked and placed in cages. A visual health inspectionwas performed on each animal to include evaluation of the coat,extremities and orifices. Each animal was also examined for any abnormalsigns in posture or movement. All animals were found to be in goodhealth and were placed on study.

The rats were acclimated for a minimum of two days prior to thecommencement of the experimental procedures, with the exception ofrandomization body weights which were collected the day followingarrival. The animals were housed individually in clear polycarbonateconventional cages or clear polycarbonate microisolator cages withcertified irradiated contact bedding. Food and water were provided adlibitum. Environmental controls were set to maintain temperatures of 18°to 26° C. (64° to 79° F.) with a relative humidity of 30% to 70%. A12:12 hour light:dark cycle was maintained.

Rats were tested for baseline threshold using the von Frey filaments onDays −4 or −1 of acclimation.

On Day 0, animals underwent a spinal nerve ligation procedure. Allsurgeries were performed under aseptic conditions. Prior to surgeries,the rats were anesthetized. The back region was shaved and prepared foraseptic surgery. The rats were placed in ventral recumbence and anincision was made just left of midline at the L4-S2 region. The leftparaspinal muscles were separated from the spinous processes (L4-S2).The L6-51 facet joint were nipped and the transverse process gentlytrimmed to provide space to access theL4 & L5 spinal nerve. The left L5and L6 spinal nerves were isolated and ligated with 6.0 silk sutures.The incision was then closed with appropriate suture material and skinwound clips. Post-operatively, Lactated Ringer's Solution (3.0-5.0 mL)was administered via subcutaneous injection to the animals.

All animals in Groups 1 and 2 received a von Frey test on Days −4 or −1,13, 14, and 17. The measurement on Day 13 was taken pre-dose. The vonFrey test for mechanical allodynia assesses anti-nociceptive propertiesof analgesic compounds. In this test, animals were first habituated tothe testing chamber so they were calm enough for their pain threshold tobe assessed. A technician blind to the treatment groups applied lightpressure to the left hind paw of the rat using a series of graded nylonfilaments (von Frey filaments) of increasing diameter. The filamentswere pressed perpendicularly against the ventral surface of the pawuntil they bent. When considered painful, the rat responds bywithdrawing its paw. Threshold allodynia was determined using theChaplan up-down method (Chaplan et al., J Neurosci Methods, 53:55-63,1994), which provides the precise force for withdrawal for each ratusing a psychophysical scale of testing.

Animals were allocated into two treatment groups on Day 13, based on vonFrey scores. Any animal that had a von Frey score greater than 6 g wasexcluded from the study. The mean von Frey scores for each group werereviewed to ensure that the mean values and standard deviation satisfiedthe assumption of homogeneity. Doses were administered by IP injectiononce on Day 13 (13 days after surgery) for Group 1 (Ab2) and Group 2(negative control antibody) (11 animals in each group; Ab2 and negativecontrol antibodies were administered at 10 mg/kg). Group 1 received anadditional IV bolus (un-anesthetized) injection of Ab2 on Day 17 priorto behavioral testing.

Blood samples for plasma were taken on Day 17 for Group 1 and analyzedfor Ab2 titer.

Outside of the expected surgical site observations and paw draggingassociated with the Chung surgery, no abnormal observations weredocumented. Treatment appeared to have no adverse effect on overallanimal health nor did it disrupt the normal weight gain expected in ratsthis age.

Results

All animals that underwent baseline testing prior to surgery on Day 0had a von Frey score of 15 (not shown) indicating normal sensitivity. OnDay 13 (prior to antibody administration), all animals had von Freyscores lower than 6 g, indicating that sensitivity to externalmechanical stimuli had developed, except for two animals which wereremoved from the study. Average von Frey scores at day 13 were less than3 g (FIG. 42, left group of bars). Following testing on day 13, animalswere administered Ab2 or a negative control antibody (10 mg/kg). On days14 and 17, von Frey scores were again tested and were higher in theAb2-treated animals than controls (FIG. 42, middle group of bars andright group of bars, respectively).

These results indicate that treatment with an anti-CGRP antibody such asAb2 may help prevent or alleviate neuropathic pain.

Example 10 First Experiment Assessing Effect of Anti-CGRP AntibodyAdministration on Analgesia (Tail Flick Model)

Three different experiments (Examples 10-12) were conducted to assessthe potential efficacy of an anti-CGRP antibody administration onanalgesia or pain. In all of these experiments a rodent tail flick (alsoreferred to as tail withdrawal) response model was used as the rodenttail flick response to radiant heat is a commonly used model to detectpotentially useful analgesic agents. This assay is particularly usefulto discriminate between centrally acting morphine-like analgesics(active) and non-opioid or peripherally acting anti-inflammatory agents(inactive). This animal model and methods and materials used therein aredescribed below.

Materials and Methods

Animals: Male Sprague Dawley derived male rats weighing 150±20 g.

Test CGRP Antibody: Ab2

Vehicle: 15 mM Histidine 250 mM Sorbitol, pH 5.5

Analgesic Compound: Morphine

Tail Flick Response Procedures: The time (seconds) required to elicit atail flick response induced by focused radiant heat was measured as thepain threshold in groups of 10 Sprague Dawley derived male rats weighing150±20 g. Baseline testing for the tail flick response was done on Day0. The rats that have a tail flick response of 3-5 seconds were includedin the study and assigned to balanced treatment groups based on baselinetail flick responses. A 15 second cut was used to avoid tissue damage.

Development of Morphine Tolerance

Each of 3 groups of 10 Male Sprague Dawley rats were dosed 2× daily viai.p. administration with saline vehicle (2 ml/kg) in the morning andevening. One of the 3 groups was in addition administered i.p. analgesic(morphine) at a dosage of 5 mg/kg 2× daily for 7 consecutive days. Asecond of the 3 groups of rats was administered i.p. an anti-CGRPantibody according to the invention (Ab2) at a dosage of 10 mg/kg as asingle bolus on day 0. The rats in the different groups were then eachtested for tail flick response once per day 30 min after the morningdose.

A one-way ANOVA followed by Dunnett's t-test is applied for comparisonbetween the vehicle control and test-compound treated groups. P<0.05 isconsidered significant.

The results of these experiments are shown in FIG. 43. The resultstherein indicate that the test CGRP antibody when administered at 10mg/kg elicited significant long-lasting analgesic effect to a thermalpain stimulus. Terminal blood samples were taken from all the testedrats via cardiac puncture and later analyzed for Ab2 titer.

Example 11 Second Tail Flick Experiment Assessing Effect of-CGRPAntibody on Analgesia) (Antibody Dose Titration)

A second set of tail flick experiments were conducted to assess theeffects of different anti-CGRP antibody dosages on analgesia using ananti-CGRP antibody according to the invention (Ab2). The rats used inthese experiments are the same type as in the previous experiment andthe tail flick protocol substantially the same. In this experimentanalgesia was compared in different groups of animals administereddifferent anti-CGRP antibody dosages in order to assess whether thedosage has an effect on analgesia. In the second set of experiments,five groups of test animals were compared as follows. A first controlgroup of animals were each administered the vehicle alone (15 mMHistidine 250 mM Sorbitol, pH 5.5), 3 groups of animals were eachadministered different dosages of the same anti-CGRP antibody containedin the vehicle (Ab2, respectively administered at dosages of 1 mg/kg, 3mg/kg or 10 mg/kg on Day 0), and a fifth group of animals wasadministered 10 mg/kg of a negative control antibody (anti-digitoxinantibody) also on Day 0.

The tail flick protocols were otherwise substantially effected asabove-described. The results were again assessed using one-way ANOVAfollowed by Dunnett's t-test for comparison between the vehicle control,negative control antibody and test-CGRP antibody treated groups. P<0.05is considered significant.

The results of these experiments are shown in FIG. 44. It can be seentherefrom that the higher antibody dosages of the test compound(inventive Ab2 anti-CGRP antibody) elicited better analgesic effectsthan the lower dosages. As anticipated the negative control antibody didnot elicit a perceptible effect on analgesia relative to the controlgroups.

Example 12 Third Tail Flick Experiment Assessing Effect of Anti-CGRPAntibody/Morphine Co-Administration on Analgesia

A third set of tail flick experiments were also conducted to assess theeffects of anti-CGRP antibody/morphine co-administration on analgesia.In these experiments a first group of animals was administered the samevehicle alone at a dosage of 5 ml/kg. A second group of animals wasadministered morphine on days 1-10 at a dosage of 5 mg/kg, administeredtwice daily, wherein such animals were on Day 0 were also administeredthe anti-CGRP antibody Ab2 at a dosage of 10 mg/kg. A third group ofanimals was administered morphine on only days 1-4 again at a dosageconcentration of 5 mg/kg, administered twice daily, and were furtheradministered the Ab2 antibody on Day 0, at a dosage of 10 mg/kg. Alladministrations were i.p.

Tail flick experiments were effected in each of these groups of animalsdaily from Day 0-10. The results of these tail flick experiments wereagain assessed using one-way ANOVA followed by Dunnett's t-test forcomparison between the vehicle control, negative control antibody andthe test anti-CGRP antibody treated group. P<0.05 is consideredsignificant.

The results of these comparisons are summarized in FIG. 45. TheAb2-treated animals receiving a daily dose of morphine throughout theexperiment exhibited morphine tolerance, and after day 5 the tail flicktime had decreased almost to the level of vehicle-treated controlanimals. In contrast, in Ab2-treated animals receiving morphine onlyuntil day 4, the tail flick time improved on day 5 and remained improveduntil day 8. The results suggest that the administration of an anti-CGRPantibody may have analgesic effects even after onset of morphinetolerance, which may be more pronounced upon withdrawal of morphine.

Example 13 Relief of Visceral Pain in Rats

Patients suffering from irritable bowel syndrome (IBS) demonstrate alower visceral sensory threshold to colorectal balloon distension(Ritchie, Gut, 1973, 14:125-32). It has been suggested in IBS that thereis heightened pain sensitivity of the brain-gut axis, with a normalpattern of activation. It has previously been shown that injection oftrinitrobenzene sulfonic acid (TNBS) into the proximal colon provokedchronic colonic hypersensitivity, measured in conscious rats by adecreased pain threshold in response to colonic distension (Diop et al.,J. Pharmacol. Exp. Ther., 2002, 302:1013-22). This chronichypersensitivity was found in the distal non-inflamed colon andpersisted for 21 days. It mimicked certain characteristics of IBS and soit can be used as a model to experimentally explore thepathophysiological aspects of this disorder. This assay is used todetermine the potential antihypersensitive effects of compounds forTNBS-induced colonic hypersensitivity.

Several studies have implicated CGRP in visceral pain (Friese et al.,Regul Pept 1997; 70:1-7; Gschossmann et al., Neurogastroenterol Motil2001; 13:229-36; Julia and Bueno, Am J Physiol 1997; 272:G141-6; Plourdeet al., Am J Physiol 1997; 273:G191-6). CGRP is the most abundantpeptide of capsaicin sensitive afferent fibers of gastrointestinalorigin, accounting for up to 80% of overall peptide immunoreactivity(Clague et al., Neurosci Lett 1985; 56:63-8; Stermini et al.,Gastroenterology 1987; 93:852-62). Additionally, injection of CGRPinduces colonic hypersensitivity in a TNBS model (Delafoy et al., 2006,Gut 55:940-5), which is reversed by a CGRP antagonist peptide (CGRP8-37).

This example describes testing of an anti-CGRP antibody in a model ofvisceral pain (TNBS-induced chronic colonic hypersensitivity) in rats.

Methods

Male Sprague-Dawley rats, weighing 390 to 450 g the day of surgery wereincluded in this study. They were housed in a temperature (19.5°C.-24.5° C.) and relative humidity (45-65%) controlled room with a 12h—light/dark cycle. Animals were housed 2 or 3 per cage and anacclimation period (at least 5 days) was observed before testing. Eachrat was identified by tail markings. The study was performed accordingto the guidelines of the Committee for Research and Ethical Issue of theI.A.S.P. (1983) and the European guidelines 2010/63/UE.

Colonic sensitivity was induced by surgical administration ofTrinitrobenzene sulfonic acid (TNBS, 50 mg/kg) 7 days before behavioraltesting. Fasted (24 hours) animals underwent surgery. Briefly, underanesthesia (Acepromazine 5 mg/kg/Ketamine 30 mg/kg), injection of TNBS(50 mg/kg, 1 ml/kg) was performed into the proximal part of the colon (1cm from the caecum). After surgery, animals were returned to their cagesin a regulated environment, and fed ad libitum until the testing day, 7days later. “Naïve” animals (rats without surgery) were placed in thesame housing conditions.

Animals were administered the anti-CGRP antibody Ab2 or a negativecontrol antibody (both at 10 mg/kg) intravenously 24 hours prior todetermination of colonic threshold. Three groups of rats were includedin this study:

Group 1: A “Naïve” group composed of animals that did not undergosurgery or TNBS treatment on D-7 and were treated with the controlantibody 24 hrs prior (i.e. D-1) to testing (i.e. measurements of thecolonic distention threshold on D0) (n=7).

Group 2: A “TNBS” group composed of animals that underwent surgery onD-7 and were treated with control antibody (24 hrs prior (i.e. D-1) totesting (i.e. measurements of the colonic distention threshold on D0)(n=8).

Group 3: A “Treated” group composed of animals that underwent surgery onD-7 and were treated with Ab2 24 hrs prior (i.e. D-1) to testing day(i.e. measurements of the colonic distention threshold on D0) (n=8).

Seven days (D7) after TNBS injection, colonic sensitivity was assessedby measuring the intra-colonic pressure required to induce a behavioralresponse during colonic distension due to the inflation of a balloonintroduced in the colon. The tests were conducted by a blindedexperimenter. This response is characterized by an elevation of the hindpart of the animal body and a clearly visible abdominal contractioncorresponding to severe contractions (Al Chaer et al., Gastroenterology2000, 119:1276-1285) and used as a pain marker (Bourdu et al.,Gastroenterology. 2005:128, 1996-2008). The balloon (5 cm) was insertedintrarectally in a minimally invasive manner to 10 cm from the anus offasted (24 h) vigil animals, and the catheter was taped to the base ofthe tail. Rats were then placed in the middle of a plexiglass box andthe catheter was connected to an electronic barostat apparatus. After a30 min-acclimation period with the inserted balloon, colonic pressurewas gradually increased by 5 mmHg steps every 30 sec from 5 to 75 mmHg(cut off) until pain behavior is evidenced. Four determinations wereperformed, 30 min, 50 min, 70 min and 90 min after balloon insertion.

Using the data from each test, the percentage of activity on colonichypersensitivity induced by the intracolonic administration of TNBS wascalculated as follows

$\left( {{Activity}\mspace{14mu} {percentage}} \right)_{Treated} = {\frac{{Distentionthreshold}_{Treated} - {Distentionthreshold}_{TNBS}}{{Distentionthreshold}_{Naive} - {Distentionthreshold}_{TNBS}} \times 100}$

Distention threshold_(Treated) is the arithmetic mean of the values forthe “Treated” group; Distention threshold_(TNBS) is the arithmetic meanof the values for the “TNBS” group; and Distention threshold_(Naive) isthe arithmetic mean of the values for the “Naïve” group.

Results

The ability of an anti-CGRP antibody to alleviate visceral pain wastested in a rat model in which chronic colonic hypersensitivity wasinduced by administration of TNBS. Visceral pain was quantified bymeasuring the colonic distension threshold, i.e., the amount ofabdominal pressure that the animals could tolerate before exhibiting abehavioral response (muscle contraction). Higher colonic distensionthreshold values indicate less sensitivity. As expected, TNBS treatmentresulted in greatly decreased the colonic distension threshold comparedto naïve animals (FIG. 46, compare middle bar (TNBS treated) and leftbar (naïve)). Ab2 administration improved the colonic distensionthreshold compared to control animals (FIG. 46, compare right bar (Ab2treated) and middle bar (control)). The improvement from Ab2administration was statistically significant (p<0.05 Student's t-test,comparison to TNBS+Negative control group). The antihypersensitiveactivity of Ab2 was computed to be 27% (indicative of the degree ofrelief of the TNBS-induced hypersensitivity).

These results suggest that anti-CGRP antibodies may be useful inpreventing or alleviating visceral pain.

1. An anti-human CGRP antibody or antibody fragment which specificallybinds to the same or overlapping linear or conformational epitope(s)and/or competes for binding to the same or overlapping linear orconformational epitope(s) on an intact CGRP polypeptide or fragmentthereof as an anti-human CGRP antibody selected from Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13 or Ab14.
 2. (canceled)3. The antibody fragment of claim 1, wherein said fragment is selectedfrom a Fab fragment, a Fab′ fragment, a F(ab′)2 fragment, or amonovalent antibody such as “metMab”.
 4. (canceled)
 5. An anti-humanCGRP antibody or antibody fragment according to claim 1, that comprisesthe same CDRs as an anti-human CGRP antibody selected from Ab1, Ab2,Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13 or Ab14. 6.The Fab fragment of claim 4, comprising a variable light chaincomprising the CDR 1 sequence of SEQ ID NO:25, the CDR 2 sequence of SEQID NO:26, and the CDR 3 sequence of SEQ ID NO:27, and/or a variableheavy chain comprising the CDR 1 sequence of SEQ ID NO:28, the CDR 2sequence of SEQ ID NO:29, and the CDR 3 sequence of SEQ ID NO:30.
 7. TheFab fragment of claim 4, comprising a variable light chain comprisingthe CDR 1 sequence of SEQ ID NO:55, the CDR 2 sequence of SEQ ID NO:56,and the CDR 3 sequence of SEQ ID NO:57, and/or a variable heavy chaincomprising the CDR 1 sequence of SEQ ID NO:58, the CDR 2 sequence of SEQID NO:59, and the CDR 3 sequence of SEQ ID NO:60.
 8. The anti-human CGRPantibody or antibody fragment of claim 1 which comprises at least 2complementarity determining regions (CDRs) in each of the variable lightand the variable heavy regions which are identical to those contained inan anti-human CGRP antibody selected from Ab1, Ab2, Ab3, Ab4, Ab5, Ab6,Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13 or Ab14.
 9. (canceled)
 10. Theanti-human CGRP antibody or antibody fragment of claim 1 which isaglycosylated or if glycosylated only contains only mannose residues.11. The anti-human CGRP antibody or antibody fragment of claim 1 whichis not N-glycosylated.
 12. The anti-human CGRP antibody or antibodyfragment of claim 1 which contains an Fc region that has been modifiedto alter effector function, half-life, proteolysis, and/orglycosylation.
 13. The anti-human CGRP antibody or antibody fragment ofclaim 1 which is a humanized, single chain or chimeric antibody.
 14. Theanti-human CGRP antibody or antibody fragment of claim 1 whichspecifically binds to CGRP expressing human cells and/or to circulatingsoluble CGRP molecules in vivo.
 15. The anti-human CGRP antibody orantibody fragment of claim 14 which specifically binds to CGRP expressedon or by human cells in a patient with a disease associated with cellsthat bind CGRP.
 16. (canceled)
 17. (canceled)
 18. (canceled) 19.(canceled)
 20. (canceled)
 21. (canceled)
 22. The anti-human CGRPantibody or fragment of claim 1 which is directly or indirectly attachedto a detectable label or therapeutic agent.
 23. A nucleic acid sequenceor nucleic acid sequences which result in the expression of ananti-human CGRP antibody or antibody fragment according to claim
 1. 24.(canceled)
 25. A vector comprising a nucleic acid sequence or sequencesaccording to claim
 23. 26. (canceled)
 27. A cultured or recombinant cellwhich expresses an antibody or antibody fragment according to any one ofclaim
 1. 28. (canceled)
 29. (canceled)
 30. The cell of claim 27 which isa diploidal Pichia yeast cell.
 31. (canceled)
 32. A method of treatmentcomprising administering to a patient with a disease or conditiontreatable by the administration of a CGRP antagonist a therapeuticallyeffective amount of at least one anti-human CGRP antibody or fragmentaccording to claim
 1. 33. The method of claim 32, wherein the disease orcondition is one associated with CGRP expressing cells a therapeuticallyeffective amount of at least one anti-human CGRP antibody or fragmentaccording to claim
 1. 34. The method of claim 32 wherein the disease isselected from migraine (with or without aura), weight loss, cancer ortumors, angiogenesis associated with cancer or tumor growth,angiogenesis associated with cancer or tumor survival, migraine, chronicmigraine, frequent episodic migraines, or menstrual migraines hemiplagicmigraines, cluster headaches, migrainous neuralgia, chronic headaches,tension headaches, general headaches, hot flushes, chronic paroxysomalhemicrania, secondary headaches due to an underlying structural problemin the head or neck, cranial neuralgia, sinus headaches (such as forexample associated with sinusitis), allergy-induced headaches ormigraines, pain, inflammatory pain, post-operative incision pain,complex regional pain syndrome, cancer pain, primary or metastatic bonecancer pain, fracture pain, osteoporotic fracture pain, pain resultingfrom burn, osteoporosis, gout joint pain, pain associated with sicklecell crises, and other nociceptic pain, as well as hepatocellularcarcinoma, breast cancer, liver cirrhosis, neurogenic pain, neuropathicpain, nociceptic pain, trigeminal neuralgia, post-herpetic neuralgia,phantom limb pain, fibromyalgia, menstrual pain, ovarialgia, reflexsympathetic dystrophy, neurogenic pain, osteoarthritis or rheumatoidarthritis pain, lower back pain, diabetic neuropathy, sciatica, orvisceral pain associated with gastro-esophageal reflux, dyspepsia,irritable bowel syndrome, inflammatory bowel disease, Crohn's disease,ileitis, ulcerative colitis, renal colic, dysmenorrhea, cystitis,menstrual period, labor, menopause, prostatitis, or pancreatitis orcomprises overactive bladder; urinary incontinence; pain; chronic pain;neurogenic inflammation and inflammatory pain; neuropathic pain; eyepain; tooth pain; post-surgical pain, trauma related pain, diabetes;non-insulin dependent diabetes mellitus and other inflammatoryautoimmune disorders, vascular disorders; inflammation; arthritis;bronchial hyperreactivity, asthma; shock; sepsis; opiate withdrawalsyndrome: morphine tolerance; hot flashes in men and women; allergicdermatitis; psoriasis; encephalitis: brain trauma; epilepsy:neurodegenerative diseases; skin diseases including pruritis, neurogeniccutaneous redness, skin rosaceousness and erythema: inflammatory boweldisease, irritable bowel syndrome, cystitis; and dysmennorhea. 35.(canceled)
 36. The method of claim 32 wherein the disease or conditionis pain, overactive bladder, urinary incontinence, headache or migraine.37. The method of claim 32, wherein the treatment further includes theadministration of another therapeutic agent or regimen selected fromanti-histamines, anti-inflammatory agents, analgesics or antibiotics.38. (canceled)
 39. (canceled)
 40. (canceled)
 41. (canceled)
 42. A methodof in vivo imaging which detects the presence of cells which expressCGRP comprising administering a diagnostically effective amount of atleast one anti-human CGRP antibody or antibody fragment according toclaim
 1. 43. The method of claim 42 wherein said administration furtherincludes the administration of a radionuclide or fluorophore thatfacilitates detection of the antibody at CGRP expressing disease sites.44. The method of claim 42 wherein the results are used to facilitatedesign of an appropriate therapeutic regimen.
 45. An isolated anti-CGRPantibody or antibody fragment comprising a V_(H) polypeptide sequenceselected from: SEQ ID NO: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103,113, 123, or 133, or a variant thereof that exhibits at least 90%sequence identity therewith; and/or a V_(L) polypeptide sequenceselected from: SEQ ID NO: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101,111, 121 or 131, or a variant thereof that exhibits at least 90%sequence identity therewith, wherein one or more of the framework (FR)or CDR residues in said V_(H) or V_(L) polypeptide has been substitutedwith another amino acid residue resulting in an anti-CGRP antibody thatspecifically binds CGRP.
 46. (canceled)
 47. (canceled)
 48. The isolatedantibody antibody or antibody fragment of claim 45 wherein at most 1 or2 of the residues in the CDRs of said V_(L) and/or the V_(H) polypeptidesequence are modified.
 49. (canceled)
 50. The antibody or antibodyfragment of claim 45, wherein said antibody is humanized.
 51. (canceled)52. (canceled)
 53. The antibody of claim 50, wherein said humanized orchimeric antibody comprises a human F_(c).
 54. The antibody or antibodyfragment of claim 53, wherein said human F_(c) is derived from IgG1,IgG2, IgG3, or IgG4.
 55. The antibody or antibody fragment of claim 45,wherein said antibody inhibits the association of CGRP with CGRP-Rand/or multimers thereof, one or more additional proteins in aCGRP-CGRP-R complex, and/or antagonizes the biological effects thereof.56. An isolated anti-CGRP antibody or antibody fragment comprising apolypeptide sequence having at least 90% or greater homology to any oneof the polypeptide sequences of claim
 45. 57. (canceled)
 58. Theantibody or antibody fragment of claim 56, wherein said antibody bindsto CGRP with an off-rate (K_(off)) of less than or equal to 10⁻⁴ S⁻¹,5×10⁻⁵ S⁻¹, 10⁻⁵ S⁻¹, 5×10⁻⁶ S⁻¹, 10⁻⁶ S⁻¹, 5×10⁻⁷ S⁻¹, or 10⁻⁷ S⁻¹. 59.The antibody or antibody fragment of claim 45, wherein said antibodyinhibits the production of a complex of CGRP with CGRP-R and/ormultimers thereof, and the production of CGRP with CGRP-R and one ormore additional proteins in a complex.
 60. An antibody fragmentaccording to of claim 45, wherein said fragment is selected from a Fabfragment, a Fab′ fragment, or a F(ab′)2 fragment.
 61. The antibody orantibody fragment of claim 45, wherein said antibody or antibodyfragment further comprises an effector moiety.
 62. The antibody orantibody fragment of claim 61, wherein the effector moiety is adetectable moiety or a functional moiety.
 63. (canceled)
 64. (canceled)65. A method of ameliorating or reducing symptoms of a disease ordisorder treatable by the administration of a CGRP antagonist comprisingadministering to an individual in need thereof a therapeuticallyeffective amount of a CGRP antibody or antibody fragment according toclaim
 45. 66. The method of claim 65, wherein the disease is associatedwith increased CGRP.
 67. The method of claim 65, wherein said disease ordisorder is selected from migraine (with or without aura), weight loss,cancer or tumors, angiogenesis associated with cancer or tumor growth,angiogenesis associated with cancer or tumor survival, hemiplagicmigraines, cluster headaches, migrainous neuralgia, chronic headaches,tension headaches, general headaches, hot flushes, chronic paroxysomalhemicrania, secondary headaches due to an underlying structural problemin the head or neck, cranial neuralgia, sinus headaches (such as forexample associated with sinusitis), allergy-induced headaches ormigraines, pain, inflammatory pain, post-operative incision pain,complex regional pain syndrome, cancer pain, primary or metastatic bonecancer pain, fracture pain, osteoporotic fracture pain, pain resultingfrom burn, osteoporosis, gout joint pain, pain associated with sicklecell crises, and other nociceptic pain, as well as hepatocellularcarcinoma, breast cancer, liver cirrhosis, neurogenic pain, neuropathicpain, nociceptic pain, trigeminal neuralgia, post-herpetic neuralgia,phantom limb pain, fibromyalgia, menstrual pain, ovarialgia, reflexsympathetic dystrophy, neurogenic pain, osteoarthritis or rheumatoidarthritis pain, lower back pain, diabetic neuropathy, sciatica, orvisceral pain associated with gastro-esophageal reflux, dyspepsia,irritable bowel syndrome, inflammatory bowel disease, Crohn's disease,ileitis, ulcerative colitis, renal colic, dysmenorrhea, cystitis,menstrual period, labor, menopause, prostatitis, or pancreatitis. 68.(canceled)
 69. The method of claim 65 wherein the disease or conditionis pain, overactive bladder, headache, or migraine.
 70. The method ofclaim 65, wherein the CGRP antibody or antibody fragment comprises thesame CDRs as Ab3 or Ab6.
 71. The method of claim 65, wherein the CGRPantibody or antibody fragment is administered in a therapeutic regimenfor treatment of a specific disease or condition associated with painthat includes the administration of another therapeutic agent.
 72. Themethod of claim 71, wherein the other therapeutic agent is selected froma chemotherapeutic, an analgesic, an anti-inflammatory, animmunosuppressant, a cytokine, an antiproliferative, an antiemetic and acytotoxin.
 73. (canceled)
 74. (canceled)
 75. (canceled)
 76. (canceled)77. (canceled)
 78. (canceled)
 79. (canceled)
 80. The method of claim 72,wherein the other active is an opioid analgesic and the combinedadministration of the opioid analgesic and the CGRP antibody or antibodyfragment increase the analgesic effect elicited thereby and/oralleviates tolerance to the analgesic.
 81. (canceled)
 82. A method ofmaking the antibody or antibody fragment of claim 45 in a polyploidyeast culture that stably expresses and secretes into the culture mediumat least 10-25 mg/liter of said antibody, comprising: (i) introducing atleast one expression vector containing one or more heterologouspolynucleotides encoding said antibody operably linked to a promoter anda signal sequence into a haploid yeast cell; (ii) producing by mating orspheroplast fusion a polyploidal yeast from said first and/or secondhaploid yeast cell; (iii) selecting polyploidal yeast cells that stablyexpress said antibody; and (iv) producing stable polyploidal yeastcultures from said polyploidal yeast cells that stably express saidantibody into the culture medium.
 83. The method of claim 82, whereinsaid yeast genera is Pichia.
 84. (canceled)
 85. (canceled)
 86. Anisolated polynucleotide comprising a polynucleotide encoding ananti-CGRP V_(H) antibody amino acid sequence selected from SEQ ID NO: 3,13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123 or 133, or encoding avariant thereof which exhibits at least 90% sequence identity therewith.87. (canceled)
 88. A vector or host cell comprising the polynucleotidesequence of claim
 86. 89. The host cell of claim 88, wherein said hostcell is a yeast cell belonging to the genus Pichia.
 90. The method ofclaim 82, wherein said heterologous polynucleotide further comprises apolynucleotide sequence encoding an anti-CGRP V_(L) amino acid sequenceselected from SEQ ID NO: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101,111, 121 or 131, or a polynucleotide encoding a variant thereof whichexhibits at least 90% sequence identity therewith.
 91. (canceled)
 92. Anisolated polynucleotide comprising the polynucleotide sequence encodingan anti-CGRP V_(L) antibody amino acid sequence selected from SEQ ID NO:1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121 or 131, or encodinga variant which exhibits at least 90% sequence identity therewith. 93.(canceled)
 94. A vector comprising the polynucleotide sequence of claim92.
 95. A host cell comprising the vector of claim
 94. 96. The host cellof claim 95, wherein said host cell is a yeast cell belonging to thegenus Pichia.
 97. The method of claim 90, wherein said heterologouspolynucleotide comprises a sequence encoding the polypeptides containedin SEQ ID NO:1 and SEQ ID NO:3; SEQ ID NO:11 and SEQ ID NO:13; SEQ IDNO:21 and SEQ ID NO:23; SEQ ID NO:31 and SEQ ID NO:33; SEQ ID NO:41 andSEQ ID NO:43; SEQ ID NO:51 and SEQ ID NO:53, SEQ ID NO:61 and SEQ IDNO:63; SEQ ID NO:71 and SEQ ID NO:73; SEQ ID NO:81 and SEQ ID NO:83; SEQID NO:91 and SEQ ID NO:93; SEQ ID NO:101 and SEQ ID NO:103; SEQ IDNO:111 and SEQ ID NO:113; SEQ ID NO:121 and SEQ ID NO:123; or SEQ IDNO:131 and SEQ ID NO:133.
 98. An anti-CGRP antibody or antibody fragmentthat binds with the same or overlapping CGRP epitope and/or competeswith an anti-CGRP antibody for binding to CGRP as an antibody orantibody fragment containing a sequence encoded by one of thepolynucleotide sequences recited in claim
 92. 99. The anti-CGRP antibodyor antibody fragment of claim 98 which binds to the same or overlappingCGRP epitope as an anti-CGRP antibody selected from Ab1, Ab2, Ab3, Ab4,Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13 or Ab14.
 100. Anisolated anti-CGRP antibody or antibody fragment comprising one or moreof the CDRs contained in the V_(H) polypeptide sequences selected from:SEQ ID NO: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, or 133and/or one or more of the CDRs contained in the V_(L) polypeptidesequences selected from: SEQ ID NO: 1, 11, 21, 31, 41, 51, 61, 71, 81,91, 101, 111, 121 or
 131. 101. The isolated anti-CGRP antibody orantibody fragment of claim 100, comprising at least 2 of the CDRscontained in a V_(H) polypeptide sequences selected from: SEQ ID NO: 3,13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123, or 133 and/or atleast 2 of the CDRs contained in the V_(L) polypeptide sequencesselected from: SEQ ID NO: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101,111, 121 or
 131. 102. The isolated anti-CGRP antibody or antibodyfragment of claim 101 comprising all 3 the CDRs contained in the V_(H)polypeptide sequences selected from: SEQ ID NO: 3, 13, 23, 33, 43, 53,63, 73, 83, 93, 103, 113, 123, or 133 and/or all 3 of the CDRs containedin the V_(L) polypeptide sequences selected from: SEQ ID NO: 1, 11, 21,31, 41, 51, 61, 71, 81, 91, 101, 111, 121 or
 131. 103. The antibody orantibody fragment of claim 100, wherein said antibody or antibodyfragment is humanized.
 104. (canceled)
 105. The antibody of claim 103,wherein said chimeric antibody comprises a human F_(c).
 106. Theantibody of claim 100, which comprises a single chain antibody.
 107. Theantibody of claim 100 which comprises a monovalent antibody molecule.108. The antibody of claim 105 wherein said human F_(c) is derived fromhuman IgG1, IgG2, IgG3, or IgG4.
 109. An isolated anti-CGRP antibody orantibody fragment comprising a V_(L) or V_(H) polypeptide sequencehaving at least 90% or greater homology to any one of the polypeptidesequences recited in claim
 100. 110. The antibody or antibody fragmentof claim 109, wherein said antibody binds to CGRP with an off-rate(k_(off)) of less than or equal to 10⁻⁴ S⁻¹, 5×10⁻⁵ S⁻¹, 10⁻⁵ S⁻¹,5×10⁻⁶ S⁻¹, 10⁻⁶ S⁻¹, 5×10⁻⁷ S⁻¹, or 10⁻⁷ S⁻¹.
 111. The antibody orantibody fragment of claim 99, wherein said antibody inhibits theassociation of CGRP with CGRP-R and/or multimers thereof, or theassociation of CGRP with its receptor (CGRP-R) and one or moreadditional proteins in a complex.
 112. An antibody fragment according toclaim 99, wherein said fragment is selected from a Fab fragment, a Fab′fragment, or a F(ab′)2 fragment.
 113. The antibody or antibody fragmentof claim 100, wherein said antibody further comprises an effector orfunctional moiety.
 114. (canceled)
 115. (canceled)
 116. (canceled) 117.A method of ameliorating or reducing symptoms of a disease or disordertreatable by the administration of a CGRP antagonist, comprisingadministering to a patient in need thereof an effective amount of anantibody or antibody fragment according to claim
 100. 118. The method ofclaim 117, wherein the disease or disorder is associated with increasedCGRP.
 119. The method of claim 118, wherein said disease or disorder isselected from migraine (with or without aura), weight loss, cancer ortumors, angiogenesis associated with cancer or tumor growth,angiogenesis associated with cancer or tumor survival, hemiplagicmigraines, cluster headaches, migrainous neuralgia, chronic headaches,tension headaches, general headaches, hot flushes, chronic paroxysomalhemicrania, secondary headaches due to an underlying structural problemin the head or neck, cranial neuralgia, sinus headaches (such as forexample associated with sinusitis), allergy-induced headaches ormigraines, pain, inflammatory pain, post-operative incision pain,complex regional pain syndrome, cancer pain, primary or metastatic bonecancer pain, fracture pain, osteoporotic fracture pain, pain resultingfrom burn, osteoporosis, gout joint pain, pain associated with sicklecell crises, and other nociceptic pain, as well as hepatocellularcarcinoma, breast cancer, liver cirrhosis, neurogenic pain, neuropathicpain, nociceptic pain, trigeminal neuralgia, post-herpetic neuralgia,phantom limb pain, fibromyalgia, menstrual pain, ovarialgia, reflexsympathetic dystrophy, neurogenic pain, osteoarthritis or rheumatoidarthritis pain, lower back pain, diabetic neuropathy, sciatica, orvisceral pain associated with gastro-esophageal reflux, dyspepsia,irritable bowel syndrome, inflammatory bowel disease, Crohn's disease,ileitis, ulcerative colitis, renal colic, dysmenorrhea, cystitis,menstrual period, labor, menopause, prostatitis, or pancreatitis orcomprises overactive bladder, pain; chronic pain; neurogenicinflammation and inflammatory pain; neuropathic pain; eye pain; toothpain; post-surgical pain, trauma related pain, diabetes; non-insulindependent diabetes mellitus and other inflammatory autoimmune disorders,vascular disorders; inflammation; arthritis; bronchial hyperreactivity,asthma; shock; sepsis; opiate withdrawal syndrome; morphine tolerance;hot flashes in men and women; allergic dermatitis; psoriasis;encephalitis; brain trauma; epilepsy; neurodegenerative diseases; skindiseases including pruritis, neurogenic cutaneous redness, skinrosaceousness and erythema; inflammatory bowel disease, irritable bowelsyndrome, cystitis; and dysmennorhea.
 120. (canceled)
 121. (canceled)122. The method of claim 118, wherein the CGRP antibody or antibodyfragment is administered in a therapeutic regimen for treatment of aspecific disease or condition associated with pain that includes theadministration of another therapeutic agent.
 123. The method of claim122, wherein the other therapeutic agent is selected from achemotherapeutic, an analgesic, an anti-inflammatory, animmunosuppressant, a cytokine, an antiproliferative, an antiemetic and acytotoxin.
 124. (canceled)
 125. (canceled)
 126. (canceled) 127.(canceled)
 128. (canceled)
 129. (canceled)
 130. (canceled)
 131. Themethod of claim 122 wherein the other active is an analgesic and whereinthe combined administration of the analgesic and the CGRP antibody orantibody fragment increase the analgesic effect elicited thereby and/oralleviates tolerance to the analgesic.
 132. (canceled)
 133. A method ofmaking the antibody of claim 100 in a polyploid yeast culture thatstably expresses and secretes into the culture medium at least 10-25mg/liter of said antibody, comprising: (i) introducing at least oneexpression vector containing one or more heterologous polynucleotidesencoding said antibody operably linked to a promoter and a signalsequence into a haploid yeast cell; (ii) producing by mating orspheroplast fusion a polyploidal yeast from said first and/or secondhaploid yeast cell; (iii) selecting polyploidal yeast cells that stablyexpress said antibody; and (iv) producing stable polyploidal yeastcultures from said polyploidal yeast cells that stably express at least10-25 mg/liter of said antibody into the culture medium.
 134. The methodof claim 133, wherein said yeast is Pichia.
 135. (canceled) 136.(canceled)
 137. An isolated polynucleotide that expresses a polypeptidecontaining at least one CDR polypeptide derived from an anti-CGRPantibody wherein said expressed polypeptide alone specifically bindsCGRP or specifically binds CGRP when expressed in association withanother polynucleotide sequence that expresses a polypeptide containingat least one CDR polypeptide derived from an anti-CGRP antibody whereinsaid at least one CDR is selected from those contained in the V_(L) orV_(H) polypeptides of SEQ ID NO: 1, 3, 11, 13, 21, 23, 31, 33, 41, 43,51, 53, 61, 63, 71, 73, 81, 83, 91, 93, 101, 103, 111, 113, 121, 123,131 or
 133. 138. The isolated polynucleotide of claim 137, thatexpresses a polypeptide containing 2 or 3 of the CDR polypeptidesderived from an anti-CGRP antibody wherein said expressed polypeptidealone specifically binds CGRP or specifically binds CGRP when expressedin association with another polynucleotide sequence that expresses apolypeptide containing 2 or 3 of the CDR polypeptides derived from ananti-CGRP antibody wherein said CDR polypeptides are selected from thosecontained in the V_(L) or V_(H) polypeptides of SEQ ID NO: 1, 3, 11, 13,21, 23, 31, 33, 41, 43, 51, 53, 61, 63, 71, 73, 81, 83, 91, 93, 101,103, 111, 113, 121, 123, 131 or
 133. 139. A vector comprising at leastone of the polynucleotide sequences of claim
 137. 140. A host cellcomprising the vector of claim
 139. 141. (canceled)
 142. The method ofclaim 133, wherein said heterologous polynucleotide comprises apolynucleotide sequences encoding at least one CDR contained in a V_(L)or V_(H) polypeptide selected from SEQ ID NO: 1, 3, 11, 13, 21, 23, 31,33, 41, 43, 51, 53, 61, 63, 71, 73, 81, 83, 91, 93, 101, 103, 111, 113,121, 123, 131 or
 133. 143. A pharmaceutical or diagnostic compositioncontaining at least one CGRP antibody or fragment according to claim 1and a pharmaceutically acceptable carrier.
 144. The pharmaceutical ordiagnostic composition of claim 143 which further comprises at least onestabilizer.
 145. The pharmaceutical or diagnostic composition of claim143 which is lyophilized.
 146. (canceled)
 147. (canceled) 148.(canceled)
 149. (canceled)
 150. A method of treating a disease orcondition treatable or preventable by the administration of a CGRPantagonist comprising administering a therapeutically effective amountof a pharmaceutical composition according to claim
 143. 151. The methodof claim 150, wherein the disease is selected from migraine (with orwithout aura), weight loss, cancer or tumors, angiogenesis associatedwith cancer or tumor growth, angiogenesis associated with cancer ortumor survival, hemiplagic migraines, cluster headaches, migrainousneuralgia, chronic headaches, tension headaches, general headaches, hotflushes, chronic paroxysomal hemicrania, secondary headaches due to anunderlying structural problem in the head or neck, cranial neuralgia,sinus headaches (such as for example associated with sinusitis),allergy-induced headaches or migraines, pain, inflammatory pain,post-operative incision pain, complex regional pain syndrome, cancerpain, primary or metastatic bone cancer pain, fracture pain,osteoporotic fracture pain, pain resulting from burn, osteoporosis, goutjoint pain, pain associated with sickle cell crises, and othernociceptic pain, as well as hepatocellular carcinoma, breast cancer,liver cirrhosis, neurogenic pain, neuropathic pain, nociceptic pain,trigeminal neuralgia, post-herpetic neuralgia, phantom limb pain,fibromyalgia, menstrual pain, ovarialgia, reflex sympathetic dystrophy,neurogenic pain, osteoarthritis or rheumatoid arthritis pain, lower backpain, diabetic neuropathy, sciatica, or visceral pain associated withgastro-esophageal reflux, dyspepsia, irritable bowel syndrome,inflammatory bowel disease, Crohn's disease, ileitis, ulcerativecolitis, renal colic, dysmenorrhea, cystitis, menstrual period, labor,menopause, prostatitis, or pancreatitis or comprises overactive bladder,pain; chronic pain; neurogenic inflammation and inflammatory pain:neuropathic pain; eye pain; tooth pain; post-surgical pain, traumarelated pain, diabetes: non-insulin dependent diabetes mellitus andother inflammatory autoimmune disorders, vascular disorders;inflammation: arthritis; bronchial hyperreactivity, asthma; shock;sepsis; opiate withdrawal syndrome; morphine tolerance; hot flashes inmen and women: allergic dermatitis; psoriasis; encephalitis: braintrauma; epilepsy; neurodegenerative diseases; skin diseases includingpruritis, neurogenic cutaneous redness skin rosaceousness and erythema;inflammatory bowel disease, irritable bowel syndrome, cystitis; anddysmennorhea.
 152. (canceled)
 153. (canceled)
 154. (canceled)
 155. Themethod of claim 150, wherein the CGRP antibody or antibody fragment isadministered in a therapeutic regimen for treatment of a specificdisease or condition associated with pain that includes theadministration of another therapeutic agent.
 156. The method of claim155, wherein the other therapeutic agent is selected from achemotherapeutic, an analgesic, an anti-inflammatory, animmunosuppressant, a cytokine, an antiproliferative, an antiemetic and acytotoxin.
 157. (canceled)
 158. (canceled)
 159. (canceled) 160.(canceled)
 161. (canceled)
 162. (canceled)
 163. (canceled) 164.(canceled)
 165. (canceled)
 166. A method of in vivo imaging whichdetects the presence of cells which express CGRP comprisingadministering a diagnostically effective amount of a diagnosticcomposition according to claim
 143. 167. The method of claim 166 whichis used as part of a planning regimen for design of an effectivetreatment protocol for migraines (with or without aura), weight loss,cancer or tumors, angiogenesis associated with cancer or tumor growth,angiogenesis associated with cancer or tumor survival, hemiplagicmigraines, cluster headaches, migrainous neuralgia, chronic headaches,tension headaches, general headaches, hot flushes, chronic paroxysomalhemicrania, secondary headaches due to an underlying structural problemin the head or neck, cranial neuralgia, sinus headaches (such as forexample associated with sinusitis), allergy-induced headaches ormigraines, pain, inflammatory pain, post-operative incision pain,complex regional pain syndrome, cancer pain, primary or metastatic bonecancer pain, fracture pain, osteoporotic fracture pain, pain resultingfrom burn, osteoporosis, gout joint pain, pain associated with sicklecell crises, and other nociceptic pain, as well as hepatocellularcarcinoma, breast cancer, liver cirrhosis, neurogenic pain, neuropathicpain, nociceptic pain, trigeminal neuralgia, post-herpetic neuralgia,phantom limb pain, fibromyalgia, menstrual pain, ovarialgia, reflexsympathetic dystrophy, neurogenic pain, osteoarthritis or rheumatoidarthritis pain, lower back pain, diabetic neuropathy, sciatica, orvisceral pain associated with gastro-esophageal reflux, dyspepsia,irritable bowel syndrome, inflammatory bowel disease, Crohn's disease,ileitis, ulcerative colitis, renal colic, dysmenorrhea, cystitis,menstrual period, labor, menopause, prostatitis, or pancreatitis. 168.The method of claim 167 wherein the treatment protocol includes one ormore of anti-histamines, anti-inflammatory agents, or antibiotics. 169.(canceled)
 170. (canceled)
 171. An isolated anti-CGRP antibody orfragment that binds to CGRP and inhibits the association of CGRP withCGRP-R, and antagonizes the biological effects thereof.
 172. Theisolated anti-CGRP antibody of claim 171, wherein the antibody isselected from Ab2, Ab3, Ab5, Ab6, Ab13, or Ab14.
 173. (canceled) 174.(canceled)
 175. (canceled)
 176. (canceled)
 177. A method of treatingpain, comprising administering an effective amount of an anti-CGRPantibody or fragment thereof which inhibits the association of CGRP withCGRP-R.
 178. (canceled)
 179. (canceled)
 180. (canceled)
 181. (canceled)182. (canceled)
 183. A method of treating migraine, comprisingadministering an effective amount of an anti-CGRP antibody or fragmentthereof which inhibits the association of CGRP with CGRP-R.
 184. Themethod of claim 183, wherein the antibody is Ab2, Ab3, Ab5, Ab6, Ab13 orAb14, or the antibody fragment is derived from Ab2, Ab3, Ab5, Ab6, Ab13or Ab14.
 185. A method of treating a non-migraine headache, comprisingadministering an effective amount of an anti-CGRP antibody or fragmentthereof which inhibits the association of CGRP with CGRP-R.
 186. Themethod of claim 185, wherein the antibody is Ab2, Ab3, Ab5, Ab6, Ab13 orAb14, or the antibody fragment is derived from Ab2, Ab3, Ab5, Ab6, Ab13or Ab14.
 187. (canceled)
 188. An anti-CGRP antibody or antibody fragmenthaving binding specificity for CGRP and comprising variable light chainCDR1, CDR2, and CDR3 polypeptide sequences and variable heavy chainCDR1, CDR2, and CDR3 polypeptide sequences selected from the following:V_(L) V_(L) V_(L) V_(H) V_(H) V_(H) CDR1 CDR2 CDR3 CDR1 CDR2 CDR3 A SeqID Seq ID Seq ID Seq ID Seq ID Seq ID No: 5 No: 6 No: 7 No: 8 No: 9 No:10 B Seq ID Seq ID Seq ID Seq ID Seq ID Seq ID No: 15 No: 16 No: 17 No:18 No: 19 No: 20 C Seq ID Seq ID Seq ID Seq ID Seq ID Seq ID No: 25 No:26 No: 27 No: 28 No: 29 No: 30 D Seq ID Seq ID Seq ID Seq ID Seq ID SeqID No: 35 No: 36 No: 37 No: 38 No: 39 No: 40 E Seq ID Seq ID Seq ID SeqID Seq ID Seq ID No: 45 No: 46 No: 47 No: 48 No: 49 No: 50 F Seq ID SeqID Seq ID Seq ID Seq ID Seq ID No: 55 No: 56 No: 57 No: 58 No: 59 No: 60G Seq ID Seq ID Seq ID Seq ID Seq ID Seq ID No: 65 No: 66 No: 67 No: 68No: 69 No: 70 H Seq ID Seq ID Seq ID Seq ID Seq ID Seq ID No: 75 No: 76No: 77 No: 78 No: 79 No: 80 I Seq ID Seq ID Seq ID Seq ID Seq ID Seq IDNo: 85 No: 86 No: 87 No: 88 No: 89 No: 90 J Seq ID Seq ID Seq ID Seq IDSeq ID Seq ID No: 95 No: 96 No: 97 No: 98 No: 99 No: 100 K Seq ID Seq IDSeq ID Seq ID Seq ID Seq ID No: 105 No: 106 No: 107 No: 108 No: 109 No:110 L Seq ID Seq ID Seq ID Seq ID Seq ID Seq ID No: 115 No: 116 No: 117No: 118 No: 119 No: 120 M Seq ID Seq ID Seq ID Seq ID Seq ID Seq ID No:125 No: 126 No: 127 No: 128 No: 129 No: 130 N Seq ID Seq ID Seq ID SeqID Seq ID Seq ID No: 135 No: 136 No: 137 No: 138 No: 139 No: 140


189. The anti-CGRP antibody or antibody fragment of claim 188, whereinsaid antibody fragment is an scFv, camelbody, nanobody, IgNAR(single-chain antibodies derived from sharks), Fab, Fab′, or F(ab′)2fragment.
 190. (canceled)
 191. The anti-CGRP antibody or antibodyfragment of claim 188, comprising a variable light chain polypeptidesequence and a variable heavy chain polypeptide sequence selected fromthe following: Variable Light Variable Heavy Chain Chain A SEQ ID NO: 1SEQ ID NO: 3 B SEQ ID NO: 11 SEQ ID NO: 13 C SEQ ID NO: 21 SEQ ID NO: 23D SEQ ID NO: 31 SEQ ID NO: 33 E SEQ ID NO: 41 SEQ ID NO: 43 F SEQ ID NO:51 SEQ ID NO: 53 G SEQ ID NO: 61 SEQ ID NO: 63 H SEQ ID NO: 71 SEQ IDNO: 73 I SEQ ID NO: 81 SEQ ID NO: 83 J SEQ ID NO: 91 SEQ ID NO: 93 K SEQID NO: 101 SEQ ID NO: 103 L SEQ ID NO: 111 SEQ ID NO: 113 M SEQ ID NO:121 SEQ ID NO: 123 N SEQ ID NO: 131 SEQ ID NO: 133


192. The anti-CGRP antibody or antibody fragment of claim 188,comprising a light chain polypeptide sequence and a heavy chainpolypeptide sequence selected from the following: Light Chain HeavyChain Ab1 SEQ ID NO: 2 SEQ ID NO: 4 Ab2 SEQ ID NO: 12 SEQ ID NO: 14 Ab3SEQ ID NO: 22 SEQ ID NO: 24 Ab4 SEQ ID NO: 32 SEQ ID NO: 34 Ab5 SEQ IDNO: 42 SEQ ID NO: 44 Ab6 SEQ ID NO: 52 SEQ ID NO: 54 Ab7 SEQ ID NO: 62SEQ ID NO: 64 Ab8 SEQ ID NO: 72 SEQ ID NO: 74 Ab9 SEQ ID NO: 82 SEQ IDNO: 84 Ab10 SEQ ID NO: 92 SEQ ID NO: 94 Ab11 SEQ ID NO: 102 SEQ ID NO:104 Ab12 SEQ ID NO: 112 SEQ ID NO: 114 Ab13 SEQ ID NO: 122 SEQ ID NO:124 Ab14 SEQ ID NO: 132 SEQ ID NO: 134


193. An anti-CGRP antibody or antibody fragment according to claim 188wherein the variable light chain and variable heavy chain polypeptidesequences are each at least 90% identical to one of the variable lightchain polypeptide sequences of SEQ ID NOS: 1, 11, 21, 31, 41, 51, 61,71, 81, 91, 101, 111, 121, or 131, and one of the variable heavy chainpolypeptide sequences of SEQ ID NOS: 3, 13, 23, 33, 43, 53, 63, 73, 83,93, 103, 113, 123 or 133, respectively.
 194. (canceled)
 195. (canceled)196. (canceled)
 197. (canceled)
 198. (canceled)
 199. (canceled) 200.(canceled)
 201. (canceled)
 202. (canceled)
 203. (canceled) 204.(canceled)
 205. An anti-CGRP antibody or antibody fragment according toclaim 188, wherein the variable light chain and variable heavy chainpolypeptide sequences are each at least 95% identical to one of thevariable light chain polypeptide sequences of SEQ ID NOS: 1, 11, 21, 31,41, 51, 61, 71, 81, 91, 101, 111, 121 or 131, and one of the variableheavy chain polypeptide sequences of SEQ ID NOS: 3, 13, 23, 33, 43, 53,63, 73, 83, 93, 103, 113, 123 or 133, respectively.
 206. An anti-CGRPantibody or antibody fragment according claim 188 wherein the variablelight chain and variable heavy chain polypeptide sequences are each atleast 98% identical to one of the variable light chain polypeptidesequences of SEQ ID NOS: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101,111, 121 or 131, and one of the variable heavy chain polypeptidesequences of SEQ ID NOS: 3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103,113, 123 or 133, respectively.
 207. An anti-CGRP antibody or antibodyfragment according to claim 188 wherein the variable light chain andvariable heavy chain polypeptide sequences are each at least 99%identical to one of the variable light chain polypeptide sequences ofSEQ ID NOS: 1, 11, 21, 31, 41, 51, 61, 71, 81, 91, 101, 111, 121 or 131,and one of the variable heavy chain polypeptide sequences of SEQ ID NOS:3, 13, 23, 33, 43, 53, 63, 73, 83, 93, 103, 113, 123 or 133,respectively.
 208. (canceled)
 209. (canceled)
 210. (canceled) 211.(canceled)